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Intraovarian insertion of autologous platelet growth factors as cell-free concentrate: Fertility recovery and first unassisted conception with term delivery at age over 40

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Int J Reprod Biomed [2020] :: Background: Use of autologous platelet-rich plasma (PRP) as an ovarian treatment has not been standardized and remains controversial. Case Presentation: A 41½ year old with diminished ovarian reserve (serum AMH=0.163mg/mL) and history of 10 unsuccessful IVF cycles presented for reproductive endocrinology consult. She and her partner declined donor oocyte IVF. They both were in good general health and laboratory tests were unremarkable, except for mild thrombocytosis (platelets=386K; normal range 150K-379K) discovered in the female. Here the patient underwent intraovarian injection of fresh platelet-derived growth factor concentrate administered as an enriched cell-free substrate. Serum AMH increased by 115% within 6wks of treatment. Results: Spontaneous ovulation occurred the month after injection and serum hCG was subsequently noted at 804mIU/mL. Following an uneventful obstetrical course, a male infant was delivered at term without complication. Conclusion: This is the first description of intraovarian injection of enriched platelet-derived growth factors followed by unassisted pregnancy and livebirth. As a refinement of conventional ovarian PRP therapy, this procedure may be particularly valuable for refractory cases where prognosis for pregnancy appears especially bleak. A putative role for thrombocytosis is also viewed in parallel with mechanisms of action advanced earlier. As experience with ovarian application of autologous platelet growth factors continues, additional research will evaluate laboratory protocol/sample preparation, injection technique, and patient selection.
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International Journal of Reproductive BioMedicine
Volume 18, Issue no. 12,
Production and Hosting by Knowledge E
Case Report
Intraovarian insertion of autologous platelet
growth factors as cell-free concentrate:
Fertility recovery and rst unassisted
conception with term delivery at age over
E. Scott Sills1, 2, 3 M.D., Ph.D., Natalie S. Rickers1, 3 LVN, Samuel H. Wood2, 3
M.D. Ph.D.
1Reproductive Biology Group IVF, FertiGen CAG; San Clemente, California USA.
2Department of Obstetrics and Gynecology, Palomar Medical Center; Escondido, California USA.
3Gen 5 Fertility Center; San Diego, California USA.
Background: The use of autologous platelet-rich plasma as an ovarian treatment has
not been standardized and remains controversial.
Case Presentation: A 41½-year old woman with diminished ovarian reserve (serum anti-
Müllerian hormone = 0.163 mg/mL) and a history of 10 unsuccessful in vitro fertilization
cycles presented for reproductive endocrinology consult. She and her partner declined
donor oocyte in vitro fertilization. They were both in good general health and laboratory
tests were unremarkable, except for mild thrombocytosis (platelets = 386K; normal
range 150-379K) discovered in the female. The patient underwent intraovarian injection
of fresh platelet-derived growth factor concentrate administered as an enriched cell-
free substrate. Serum anti- Müllerian hormone increased by 115% within 6 wks of
treatment. Spontaneous ovulation occurred the month after injection and subsequently
the serum human chorionic gonadotropin was noted at 804 mIU/mL. Following an
uneventful obstetrical course, a male infant was delivered at term without complication.
Conclusion: This is the rst description of intraovarian injection of enriched platelet-
derived growth factors followed by unassisted pregnancy and live birth. As a renement
of conventional ovarian platelet-rich plasma therapy, this procedure may be particularly
valuable for refractory cases where prognosis for pregnancy appears especially bleak.
A putative role for thrombocytosis is also viewed in parallel with mechanisms of action
as advanced earlier. With continued experience in ovarian application of autologous
platelet growth factors, additional research will evaluate laboratory protocol/sample
preparation, injection technique, and patient selection.
Key words: Ovarian rejuvenation, Platelet-rich plasma, Cytokines, Infertility, IVF.
How to cite this article:Sills ES, Rickers NS, Wood SH. “Intraovarian insertion of autologous platelet growth factors as cell-free concentrate: Fertility recovery
and rst unassisted conception with term delivery at age over 40,” Int J Reprod BioMed 2020; 18: 1081–1086. Page 1081
Corresponding Author:
E. Scott Sills, P.O. Box 73910
San Clemente, California
92673 USA.
Tel: (+1) 949 -899-5686
Received 18 July 2020
Revised 9 September 2020
Accepted 8 November 2020
Production and Hosting by
Knowledge E
Sills et al. This article is
distributed under the terms
of the Creative Commons
Attribution License, which
permits unrestricted use and
redistribution provided that
the original author and source
are credited.
Aatoonian Abbas M.D.
International Journal of Reproductive BioMedicine Sills et al.
1. Introduction
It is well-known that as women age, both the
quality and the quantity of eggs decline; the low
ovarian reserve observed among older infertile
patients occurs as an expected physiological
consequence of normal ovarian senescence.
In such cases, even the use of high-dose
gonadotropin protocols is generally futile, leaving
oocyte donation/IVF as the only clinically effective
treatment (1-3).
As an investigational alternative to egg donation,
the surgical placement of autologous platelet-
rich plasma (PRP) into ovarian tissue rst began
to attract attention in 2016 (4). This pioneering
technique of ovarian “rejuvenation” was followed
by two publications describing similar use of PRP
for poor-prognosis patients as a precursor to IVF.
Specically, four patients with undetectable or
very low ovarian reserve (mean age 42 years)
who had planned for donor egg treatment instead
underwent the PRP treatment; all four patients
developed blastocysts from their oocytes (5), one
of them has since undergone thaw, transfer, and
had a healthy term delivery. Experts in Greece
also described three poor-responder IVF patients
(mean age 38 years) with similar “revolutionary”
responses (6). At least one patient who produced
only embryos with genetic errors was able to
achieve “ploidy rescue” following intraovarian
injection of platelet-derived growth factors (PDGFs)
before IVF, culminating in successful term live birth
(7). Ovarian PRP has been formally evaluated in
a descriptive pilot study including >150 patients,
where no signicant change was observed in the
serum AMH of most patients (8). However, the
measured response rate after ovarian PRP (28%)
approximates the overall IVF pregnancy rate in the
USA. While ovarian treatment with growth factors
has generally been framed as a precursor to IVF (5),
almost no data exist on the reproductive outcome
in the absence of IVF. Here, intraovarian injection of
enriched, cell-free platelet growth factors followed
by healthy term delivery - with no gonadotropins or
IVF - is presented.
2. Case Presentation
A 41½-year old woman presented with her
husband (aged 41) for reproductive endocrinology
consultation. They were both in good general
health and took no regular medication. Her
past surgical history was signicant for an
uncomplicated laparoscopic myomectomy in
2018. The assessment of endometrial cavity
contour after the procedure was normal. At age
38, the patient was provisionally diagnosed with
primary ovarian insufciency (POI) based on
repeatedly elevated levels of follicle-stimulating
hormone (FSH) and “undetectable” anti-Müllerian
hormone (AMH). The remainder of the work-
up in this case was essentially unremarkable,
including a normal endocrine prole and negative
pregnancy test, although a mild thrombocytosis
(platelets = 386K; normal range 150-379K)
was discovered in the female. Moreover, two
pregnancies were established without medical
assistance >5 yrs ago, but both were electively
terminated without complication. The couple had
initiated at least 10 IVF cycles elsewhere before
consultation, however, none were successful; chart
review attributed these failures to poor follicular
response, “empty follicles,” fertilization failure,
or culture arrest. Additional IVF attempts were
discouraged, and for personal reasons the couple
declined donor egg IVF.
Here, the patient was counseled and a written
informed consent was obtained for the injection
of enriched autologous PDGFs into both ovaries.
This treatment was offered as an extension of
a previous IRB-approved prospective clinical
trial (8). PDGFs were isolated rst via obtaining
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International Journal of Reproductive BioMedicine Fertility recovery with platelet growth factors
autologous PRP activated by calcium gluconate
as previously described (8), followed by additional
enrichment by centrifugation at 250 g ×15 min
with phosphate-buffered saline (Thermo Fisher
Scientic, Carlsbad Calif USA) irrigation of the
platelet pellet ×3. Thereafter, the resuspended
platelet pellet was next processed through
a centrifugation sequence (300 ×g for 10
min, 2,000 ×g for 10 min) to subtract debris,
based on prior protocols (9-11). The resulting
cell-free supernatant was maintained fresh at
room temperature for ovarian insertion. After
processing, a volume of approximately 1.5 mL
was injected into ovarian stroma (subcapsular)
under direct transvaginal ultrasound guidance
with instrumentation as for conventional PRP
dosing (5). The procedure was well tolerated
with no complications, completed in <10 min,
and required no anesthesia or sedation. Using a
uniform assay (12), the serum AMH was checked
again the following month, and the level had
increased from 0.163 ng/mL pretreatment to
0.352 ng/mL six weeks later (Figure 1). Ovulation
occurred without medical assistance and serum
human chorionic gonadotropin was subsequently
noted at 804 mIU/mL. Her prenatal course was
uneventful and she delivered a healthy 3,740
gr male infant without complication by elective
cesarean at 39 weeks gestation. Both the mother
and the baby continue to do well.
Figure 1. Serum AMH levels measured before versus after bilateral intraovarian injection of platelet-derived growth factors.
Approximately six weeks after the treatment (blue arrow), an unassisted pregnancy was conrmed (green arrow). Baseline
thrombocytosis is depicted at left (red arrow) to show nominally elevated pretreatment platelet concentration relative to the
expected reference range (vertical bar). AMH: Anti-mullerian hormone; INJ: Intraovarian injection of enriched platelet growth
factors; hCG: Human chorionic gonadotropin; PLT: Platelet concentration; Dashed/solid line: Estimated/veried data.
2.1. Ethical consideration
Written informed consent was obtained from the
patient, who read and approved the manuscript
before publication.
3. Discussion
For both patients and providers, the problem of
recurrent IVF failure is difcult and usually leads
to discussion about donor oocytes. Although Page 1083
International Journal of Reproductive BioMedicine Sills et al.
this approach is an established component of
advanced fertility treatment since rst introduced in
the 1980s (13, 14), it is still sometimes unacceptable
to some patients. For this reason, any advances
to open safe and effective opportunities for
patients would be welcomed. While successful
pregnancy has been reported even when POI
has been validated (15), the numerous IVF failures
experienced by our patient focused renewed
attention on intraovarian injection of autologous
platelet-derived cytokines. Although no consensus
exists regarding any preferred PRP sample
preparation or injection technique, there is even
less agreement regarding an optimal methodology
for the newer approach for incubation and/or
isolation of platelet growth factors.
As summarized in Figure 2, selected
platelet releasate components are shown with
depleted platelets (DEP). These include TGF-β,
a transcription activator modulating genes for
differentiation, chemotaxis, and proliferation and
activation of immune system cells; vascular
endothelial growth factor, a signal protein
stimulating blood vessel formation; insulin-like
growth factors (1 and 2), proteins required for
cell stimulation; PDGF, critical to blood vessel
growth from adjacent capillaries, mitogenesis,
and proliferation of mesenchymal cells including
broblasts, osteoblasts, tenocytes, vascular
SMCs, and mesenchymal stem cells; Interleukin-
1β (IL-1β), an inammatory marker involved in
cell growth, differentiation, and programmed
death; Interleukin-8 which initiates angiogenesis,
perfusion, and movement to injury/infection sites;
epidermal growth factor, a key messenger in
cell proliferation, differentiation, and survival;
as well as basic broblast growth factor, a
mediator with mitogenic and cell survival
activities including embryonic development,
cell growth, morphogenesis, and tissue
Placing such growth factors within ovarian tissue
may potentiate higher AMH output and improve IVF
response in several ways (16). One possibility is that
any follicle emerging after intraovarian injection
of these growth factors was merely latent, not
completely absent. Perhaps more controversially,
PDGFs could engage with uncommitted ovarian
stem cells (17) and work along multiple signaling
pathways to evoke differentiation to de novo
oocytes. Indeed, PRP has been shown to induce
proliferation of some cell populations to improve
stemness and to enhance in vitro expression of
receptivity markers (18, 19). Since these factors
also have angiogenic properties, it is plausible that
improving capillary ow and thus tissue oxygen
delivery might induce benecial ovarian effects
after injection (20).
While it is tempting to ascribe any effects
observed here to platelet cytokines, it is not
possible to separate this component from the
injection process itself where ovarian micro-
puncture alone might be therapeutic (21, 22).
However, we believe the link between the
treatment and subsequent pregnancy here is
supported by the brief interval between treatment
and pregnancy, the absence of any other therapy,
the serum AMH pattern after injection, and
thrombocytosis. This latter issue draws notice to
the role of platelet dynamics when ovarian PRP and
related treatments are critically assessed; ambient
platelet count has been identied as a modulator
of AMH response independent of patient age,
infertility duration, or pre-injection AMH level (5,
We agree that limitations exist whenever
case data are considered. For example,
the marked uptick in serum AMH and the
favorable reproductive outcome attained
here after treatment are associative and not
necessarily causative. Additional study should
help clarify signaling pathways involved in
Page 1084
International Journal of Reproductive BioMedicine Fertility recovery with platelet growth factors
follicular development, thereby providing potential
techniques to make pregnancy possible even
for older patients with low or absent ovarian
Figure 2. Outline comparing standard platelet-rich plasma (PRP) versus enriched platelet factors (EPF) ovarian treatment.
Conict of Interest
ESS and SHW have received a provisional U.S.
patent for the process & treatment of ovarian
disorders using platelet cytokine derivatives. NSR
has no conicts to disclose.
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Full-text available
The inverse correlation between maternal age and pregnancy rate represents a major challenge for reproductive endocrinology. The high embryo ploidy error rate in failed in vitro fertilization (IVF) cycles reflects genetic misfires accumulated by older oocytes over time. Despite the application of different follicular recruitment protocols during IVF, gonadotropin modifications are generally futile in addressing such damage. Even when additional oocytes are retrieved, quality is frequently poor. Older oocytes with serious cytoplasmic and/or chromosomal errors are often harvested from poorly perfused follicles, and ovarian vascularity and follicular oxygenation impact embryonic chromosomal competency. Because stimulation regimens exert their effects briefly and immediately before ovulation, gonadotropins alone are an ineffective antidote to long-term hypoxic pathology. In contrast, the tissue repair properties (and particularly the angiogenic effects) of platelet-rich plasma (PRP) are well known, with applications in other clinical contexts. Injection of conventional PRP and/or its components (e.g., isolated platelet-derived growth factors as a cell-free substrate) into ovarian tissue prior to IVF has been reported to improve reproductive outcomes. Any derivative neovascularity may modulate oocyte competence by increasing cellular oxygenation and/or lowering concentrations of intraovarian reactive oxygen species. We propose a mechanism to support intrastromal angiogenesis, improved follicular perfusion, and, crucially, embryo ploidy rescue. This last effect may be explained by mRNA upregulation coordinated by PRP-associated molecular signaling, as in other tissue systems. Additionally, we outline an intraovarian injection technique for platelet-derived growth factors and present this method to help minimize reliance on donor oocytes and conventional hormone replacement therapy.
Full-text available
One explanation for why downstream gonadotropin protocol changes during IVF commonly arrive too late to have significant effects is that embryo development actually begins during oogenesis. Thus, efforts to modify the chromosomal status of blastocysts must address the ovarian milieu well in advance of follicular recruitment. A 42 year old woman with primary infertility of 3 year duration attended with her partner. Five previous IVF cycles had produced 20 embryos, but all had genetic abnormalities and no embryo transfer was performed. Karyotypes and all lab tests were normal for both partners. 3 months before her IVF here, she received isolated platelet-derived growth factors injected into both ovaries as a cell-free, enriched substrate. Genetic assessments were via whole genome amplification and DNA tagmentation and PCR adapter sequences. Comprehensive chromosomal screening was carried out by dual-indexed sequencing of pooled libraries on the MiSeq™ platform. From this IVF cycle one euploid 46, XY blastocyst was produced and vitrified on the day of trophectoderm biopsy. 9 days after frozen embryo transfer, serum human chorionic gonadotropin was 250 mIU/ml and a transvaginal ultrasound at 6 week gestation confirmed a single intrauterine pregnancy with fetal heart at 153/min. A healthy male infant was delivered by c-section at 39 weeks' gestation. While cellular and molecular events directing the oocyte-to-embryo transition are incompletely characterized, processes related to ovarian stem cell differentiation, mitochondrial dynamics, and mRNA storage, translation, and degradation likely are relevant. It appears that intraovarian application of autologous platelet-derived growth factors, when used before IVF, can impact oocyte integrity and facilitate euploid blastocyst development. Although research on intraovarian injection of autologous activated platelet rich plasma has already shown improved quantitative IVF responses, this is the first description of qualitative improvements in embryo genetics after intraovarian injection of autologous platelet-derived growth factors.
Full-text available
Background: Primary ovarian insufficiency (POI) is a devastating diagnosis for reproductive-aged women due to the associated infertility and other serious health consequences. Spontaneous pregnancy without hormone replacement therapy (HRT) and/or assisted reproductive technology (ART) rarely occurs in POI patients, particularly in those patients with long-term amenorrhea. Case: On March 4, 2019, a 31-year-old Chinese POI patient visited our hospital for a spontaneous pregnancy after 10 years of amenorrhea and discontinuation of HRT 4 years prior. The patient had menarche at the age of 13, with 3 years of regular menstruation followed by amenorrhea occurring at the age of 20. POI was diagnosed by several hospitals; chromosome examination revealed a normal 46, XX karyotype. Treatment with estradiol valerate and progesterone did lead to resumed menstruation, while amenorrhea resumed after withdrawal of HRT. The patient married at the age of 23 and tried to conceive by HRT until the age of 25; her beta-human chorionic gonadotropin (HCG), estrogen (E2), and progesterone levels were 32987.7~119151.4 mIU/ml, 671.0~>1,000 pg/mL, and 6.6~27.9 ng/ml, respectively. On March 22, 2019, ultrasonography showed an intrauterine pregnancy with a normally developed gestational sac sized 45 × 42 × 32 mm with a 17 mm crown-rump length. On October 29, a 3,400 g healthy girl baby was delivered; the patient had a spontaneous delivery with natural labor. Conclusion: Spontaneous pregnancy is possible in women with POI and 10 years of amenorrhea.
Full-text available
Premature ovarian failure (POF) usually happens former to the age of 40 and affects the female physiological state premenopausal period. In this condition, ovaries stop working long before the expected menopausal time. Of diagnostic symptoms of the disease, one can mention amenorrhea and hypoestrogenism. The cause of POF in most cases is idiopathic; however, cancer therapy may also cause POF. Commonly utilized therapies such as hormone therapy, in‐vitro activation, and regenerative medicine are the most well‐known treatments for POF. Hence, these therapies may be associated with some complications. The aim of the present study is to discuss the beneficial effects of tissue engineering for fertility rehabilitation in patients with POF as a newly emerging therapy.
Full-text available
In clinical infertility practice, one intractable problem is low (or absent) ovarian reserve which in turn reflects the natural oocyte depletion associated with advancing maternal age. The number of available eggs has been generally thought to be finite and strictly limited, an entrenched and largely unchallenged tenet dating back more than 50 years. In the past decade, it has been suggested that renewable ovarian germline stem cells (GSCs) exist in adults, and that such cells may be utilized as an oocyte source for women seeking to extend fertility. Currently, the issue of whether mammalian females possess such a population of renewable GSCs remains unsettled. The topic is complex and even agreement on a definitive approach to verify the process of ‘ovarian rescue’ or ‘re-potentiation’ has been elusive. Similarities have been noted between wound healing and ovarian tissue repair following capsule rupture at ovulation. In addition, molecular signaling events which might be necessary to reverse the effects of reproductive ageing seem congruent with changes occurring in tissue injury responses elsewhere. Recently, clinical experience with such a technique based on autologous activated platelet-rich plasma (PRP) treatment of the adult human ovary has been reported. This review summarizes the present state of understanding of the interaction of platelet-derived growth factors with adult ovarian tissue, and the outcome of human reproductive potential following PRP treatment.
Full-text available
Purpose To investigate a possible influence of repetitive micro-traumata on the ovaries in the course of oocyte retrieval during IVF/ICSI treatment on serum anti-Müllerian hormone (AMH) levels. Methods The study included retrospectively collected data from women who underwent three or more consecutive IVF/ICSI treatments between 2007 and 2017. The primary endpoint of the study was to evaluate changes in serum AMH levels on cycle days 1–3 during the course of repetitive IVF/ICSI treatments. Results A total of 125 patients were included in this study. Median AMH levels before the first, second and third IVF/ICSI cycles were 3.8 ng/mL (IQR 1.8–7.1), 3.3 ng/mL (IQR 1.8–6.1) and 3.0 ng/mL (IQR 1.6–5.3), respectively (p = n.s.). In patients who underwent IVF/ICSI due to polycystic ovary syndrome (PCOS), we found a significant decrease in AMH serum levels between the first [AMH 9.7 ng/mL (IQR 7.4–14.4)] and the third [AMH 5.3 ng/mL (IQR 3.3–10.4)] IVF/ICSI cycles (p = 0.026). When performing a generalized linear model, we found PCOS to be an independent predictor for serum AMH decrease during the course of three oocyte retrievals (p < 0.001). Conclusions When comparing the indications for IVF/ICSI, we observed a significant decrease in AMH serum levels after repetitive oocyte retrievals only in women with PCOS, while the decrease in AMH was not significant in patients with tubal factor, endometriosis, male factor and unexplained infertility. This finding leads us to hypothesize that repetitive micro-traumata on the ovarian cortex might diminish/normalize functional ovarian reserve in women with PCOS. Further prospective studies are highly warranted to allow firm conclusions.
Full-text available
Background Intrauterine adhesion (IUA) is a major cause of female secondary infertility. We previously demonstrated that menstrual blood-derived stromal cell (MenSC) transplantation helped severe IUA patients have pregnancy and endometrium regeneration. We also initiated platelet-rich plasma (PRP) acted as a beneficial supplement in MenSC culturing and a potential endometrial receptivity regulator. Here, we investigated the therapeutic effect of combined transplantation of MenSCs with PRP in rat IUA models and the mechanisms of MenSCs in endometrium regeneration. Methods Rat IUA models were established by intrauterine mechanical injured. Nine days later, all rats were randomly assigned to four groups received different treatment: placebo, MenSC transplantation, PRP transplantation, and MenSCs + PRP transplantation. The traces of MenSCs were tracked with GFP label. Endometrial morphology and pathology, tissue proliferation, inflammation, pregnancy outcomes, and mechanism of MenSCs in the regeneration of endometrium were investigated. Results Notably, at days 9 and 18 post-treatment, MenSC transplantation significantly improved endometrial proliferation, angiogenesis, and morphology recovery and decreased collagen fibrosis and inflammation in the uterus. MenSCs had lesion chemotaxis, colonized around the endometrial glands. Gene expression of human-derived secretory protein IGF-1, SDF-1, and TSP-1 was detected in the uterus received MenSCs at day 18. The three treatments can all improve fertility in IUA rats. Moreover, gene expressions of cell proliferation, developmental processes, and other biological processes were induced in MenSC transplantation group. Hippo signaling pathway was the most significantly changed pathway, and the downstream factors CTGF, Wnt5a, and Gdf5 were significantly regulated in treatment groups. PRP enhanced these parameters through a synergistic effect. Conclusions In summary, MenSCs could effectively improve uterine proliferation, markedly accelerate endometrial damage repairment and promote fertility restoration in IUA rats, suggesting a paracrine restorative effect and Hippo signaling pathway stimulation. Our results indicate MenSCs, a valuable source of cells for transplantation in the treatment intrauterine adhesion. Combined with PRP, this cell therapy was more effective. Electronic supplementary material The online version of this article (10.1186/s13287-019-1155-7) contains supplementary material, which is available to authorized users.
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Study objective To compare pregnancy outcomes in PCOS women undergoing transvaginal ovarian injury (TVOI) and laparoscopic ovarian drilling (LOD) Design 126 infertile patients with PCOS were included in this prospective cohort study Canadian task force classification of level of evidence IIA. Setting University-affiliated fertility center. Patients Sixty-seven infertile patients with the history of failed in vitro maturation underwent follow-up as the TVOI group. Fifty-nine infertile women who underwent LOD acted as controls. All subjects had PCOS with menstrual irregularity and were anovulatory by repetitive serum progesterone levels. Interventions The LOD group underwent six cauterizations of a single ovary with 30W for 4–6 s. Failed IVM subjects with 20–30 needle punctures per ovary acted as the TVOI group. Subjects were followed for six months. Measurements and main results There was not a significant difference between the groups when the cases were evaluated in terms of spontaneous pregnancy or miscarriage rates. BMI levels decreased in both the TVOI and the LOD groups in a similar fashion. However, serum AMH and AFC decreased greater after LOD than they did with TVOI over the six-month duration of the study (p < 0.001 in both cases). Conclusions Preliminary data suggest that TVOI likely represents a safer, less costly and equally effective manner of surgical ovulation induction in anovulatory PCOS women when compared to LOD.
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The aim of this study was to investigate the influence of different incubation methods on the growth factor content of lysates of platelet-rich fibrin (PRF), advanced-platelet-rich fibrin (A-PRF) and platelet-rich plasma (PRP) products. A comparison of related studies suggests that the method of sample preparation has a significant influence on growth factor content. There are few reports on the comparison of non-Ca²⁺-activated PRP, Ca²⁺-activated PRP, A-PRF, and PRF, along with a lack of information on the release of PDGF-BB, TGF-β1, and VEGF among the different incubation methods. The lysate preparation was made of non-Ca²⁺-activated PRP, Ca²⁺-activated PRP, PRF, and A-PRF, using a room-temperature, 37 °C, or freeze–thaw–freeze incubation method. Afterwards the VEGF, PDGF-BB, and TGF-β1 content was investigated by running ELISA tests. Growth factor levels were significantly increased in the non-Ca²⁺-activated PRP with freeze–thaw–freeze incubation, and in the PRF preparation there was a significant disadvantage to using room temperature incubation for releasing growth factors. In conclusion, the freeze–thaw–freeze method is sufficient for releasing growth factors, and calcium activation is not necessary. Finally, the study demonstrates the possibility of preparing PRP products from platelet concentrates, so that preoperative blood sampling might not be required.