Wound healing activity of Hymenocallis littoralis - Moving beyond ornamental plant

Abstract

Spider lily (Hymenocallis littoralis) belongs to Amaryllidaceae family is a well-known plant species for its
medicinal properties. The inhibitory effects of H. littoralis methanol sonication extracts were evaluated for wound
healing activity. This is the first report on the wound healing activity of Malaysian origin H. littoralis. The bulb, flower,
root, anther, stem and leaves of H. littoralis methanol sonication extracts were used for scratch-wound assay. The cell
line was treated with two different concentrations; 1 and 10μg/ml of extracts. The extracts were prepared freshly by
dissolving in sterile phosphate saline buffer (PBS) and the healing activity was observed from 2, 4, 8, 12, 24, 36 and 48
h. The bulb, root, stem and anther methanol extracts demonstrated active wound healing activities at 1 μg mL-1at 36 h of
treatment. At the low concentration the bulb, root, stem and anther methanol extracts heals the wound compared to leaf
and flower extracts. It’s demonstrated that these extracts contain effective phytochemical substances which are
responsible for wound healing process. This finding suggests the potential application of H. littoralis methanol extract in
wound healing activity.

Full text

Pak. J. Pharm. Sci., Vol.31, No.6, November 2018, pp.2537-2543 2537
REPORT
Wound healing activity of Hymenocallis littoralis - Moving beyond
ornamental plant
Jeevandran Sundarasekar1, Geethaa Sahgal2, Vikneswaran Murugaiyah3, Lam Kit Lay2,
Ong Ming Thong2 and Sreeramanan Subramaniam1*
1School of Biological Sciences, Universiti Sains Malaysia, Minden, Penang, Malaysia
2Institute for Research in Molecular Medicine (INFORMM), Universiti Sains Malaysia, Minden, Penang, Malaysia
3School of Pharmaceutical Sciences Universiti Sains Malaysia, Minden, Penang, Malaysia
Abstract: Spider lily (Hymenocallis littoralis) belongs to Amaryllidaceae family is a well-known plant species for its
medicinal properties. The inhibitory effects of H. littoralis methanol sonication extracts were evaluated for wound
healing activity. This is the first report on the wound healing activity of Malaysian origin H. littoralis. The bulb, flower,
root, anther, stem and leaves of H. littoralis methanol sonication extracts were used for scratch-wound assay. The cell
line was treated with two different concentrations; 1 and 10µg/ml of extracts. The extracts were prepared freshly by
dissolving in sterile phosphate saline buffer (PBS) and the healing activity was observed from 2, 4, 8, 12, 24, 36 and 48
h. The bulb, root, stem and anther methanol extracts demonstrated active wound healing activities at 1 µg mL-1at 36 h of
treatment. At the low concentration the bulb, root, stem and anther methanol extracts heals the wound compared to leaf
and flower extracts. It’s demonstrated that these extracts contain effective phytochemical substances which are
responsible for wound healing process. This finding suggests the potential application of H. littoralis methanol extract in
wound healing activity.
Keywords: Hymenocallis littoralis, Methanol sonication, Root, Bulb, Hs27 cell line.
INTRODUCTION
Wound healing comprises continues cell-cell and cell-
matrix interactions in three overlapping phases such as
inflammation (0-3days), cellular proliferation (3-12 days)
and finally remodeling of the wound (3-6 months)
(Schmidt et al., 2009). A thick actin bundles of
myofibroblasts expressed in wounded dermis (Houghton
et al., 2005; Gurtner et al., 2008). Nevertheless in the
presence of oxygen free radicals, microbial infections and
UV rays could reduce the wound recovery mechanism
process (Houghton et al., 2005; Kumar et al., 2007;
Schmidt et al., 2009). Traditional herbal medicine with
effective pharmacological activities such as antioxidant
and antimicrobial often used in broad area in various skin
diseases (Suntar et al., 2010) includes cuts, wounds and
burns (Kumar et al., 2007). Numerous herbs were
scientifically proven to be used as remedies to cure the
wounds in animal based studies (Dalazen et al., 2005).
Wound healing property of Hymenocallis littoralis
(Amaryllidaceae) extract was evaluated for the first time
using cell based techniques. Hymenocallis littoralis an
ornamental plant exhibits numerous therapeutic properties
such as anti-Candida, antioxidant, cytotoxicity and wound
healing activities (Abou-Donia et al., 2008).
Fascinatingly, this exploration was carried out because
Aloe vera (Xanthornrhoeaceae) is the only plant which
was used cell cultured-based technique to evaluate the
wound healing activity (Krishnan, 2006). Currently, there
are reports on anticancer property of H. littoralis extract
but there are no reports on the wound healing activity
(Idso et al., 2000; Ingrassia et al., 2008). Thus, bulb,
anther, stem, leaves, flower and root's crude extracts were
subjected to the wound healing assessment using human
foreskin fibroblast cell line (Hs27). The human fibroblast
in vitro model is essential to correlate the contractile
events of wound (Marharet et al., 1998; Theim and
Grosslinka, 2003).
MATERIALS AND METHODS
Samples preparation
H. littoralis plants were bought from Penang Botanical
Gardens and the authenticity work was carried out by Mr.
Shanmugam (Curator) from School of Biological
Sciences, Universiti Sains Malaysia. Each of the plant
parts (leaves, stem, root, bulbs, flowers, and anther) were
cut into small pieces, washed in running tap water and
dried at 40°C in a sterile oven for a week to remove the
moisture content. The samples were powdered using
blender (Panasonic, 380V) and extracted using methanol
solvent by sonication technique to obtain the crude
extracts. The sonication was run for 5 min and the extract
*Corresponding author: e-mail: sreeramanan@gmail.com

Wound healing activity of Hymenocallis littoralis - Moving beyond ornamental plant
Pak. J. Pharm. Sci., Vol.31, No.6, November 2018, pp.2537-2543
2538
was filtered through filter paper (Whatman No.1). The
technique is repeated for three times using the residues.
The filtrate was collected and concentrated in a rotary
evaporator (RII0 Buchi, Switzerland) at 40°C. The
concentrated extract was dried in an oven at 40°C for
three days to obtain consistent weight and freeze dried for
2 days (Masoko and Eloff, 2007). The stock solution of H.
littoralis methanolic extract of flower, root, bulb, leaf,
stem and anther were prepared in 0.5% (v/v)
dimethylsulfoxide (DMSO). The working solutions for
every extracts were prepared at 1 and 10µg mL-
1concentrations via dilution technique and diluted in
complete medium
Cell line and culture conditions
The human foreskin fibroblast (Hs27; ATCC CRL-1634)
was used to study wound healing property of H. littoralis.
The cell line is gifted by Dr. Ong Ming Thong from
Institution for Research in Molecular Medicine
(INFROMM), Universiti Sains Malaysia. The stock
culture was maintained in a 25 cm2 flask in Dulbecco's
modified Eagle's medium high glucose (DMEM-H)
(GIBCO, Invitrogen, United States of America)
supplemented with 10% (volume/volume) fetal bovine
serum (FBS) (GIBCO, Invitrogen, USA) and 1% (v/v)
anti-mycotic antibiotic (GIBCO, Invitrogen, USA). The
cells were incubated at 37°C in a humidified carbon
dioxide incubator (5% CO2). Cells at passages 10 and 11
were used in this study (Yue et al., 2010).
Scratch-wound assay
Hs27 cells culture were seeded at density of 2,500 cells
per well into the 96 well flat-bottom micro plates (Nunc)
with incomplete medium (DMEM-H and 1% (v/v)
antibiotic-antimycotic) for 2 days to synchronize the cell
growth. The cells were observed daily under inverted
microscope (MOTIC, AE 31 model, USA) to confirm the
growth condition of the cells. On 48h, the cells were
wounded using sterile p-200 pipette tips (Axygen T- 300)
and the cells washed gently twice using sterile phosphate
buffer saline (PBS, pH 7). The wounded cells were treated
with 120µL of H. littoralis plant extracts with 10 and 1µg
ml-1 concentrations. The cells were observed at 0, 2, 4, 8,
12, 24, 36 and 48 h to analysis the cell migration and the
images were captured for each observation time points
(Liang et al., 2007). Cells treated with complete medium
denoted as control while cells treated with 0.05% DMSO
solvent stand as negative control. The complete medium
is consists of 5% FBS, 1% antibiotic and basal media
(DMEM-High glucose) to provide complete nutrients for
cell growth.
Image capture and data analysis
Images at time zero (t= 0) were captured to record the
initial diameter of the wounds and the recovery of the
wounded due to cell migration toward the denuded
diameter was evaluated at 2, 4, 8, 12, 24, 36 and 48 h. The
images were captured using an Apotome microscope
(Axio Vision 4, Carl Zeiss, USA). The diameter of wound
was measured by Axio Vision Release 4.8.1 software
(Carl Zeiss, USA). The migration of the cells was
expressed as percentage of wound closure:
Percentage (%) of
wound closure = %100
0h-At
Äh)-At0h -[(At ×
⎥
⎦
⎤
⎢
⎣
⎡
Where, At-0h is the length of wound measured
immediately after scratching and At-∆h is the length of
wound measured at interval time points (4, 8, 12, 24, 36,
48 hr) after scratching (Yue et al., 2010).
STATISTICAL ANALYSIS
Each experiment was performed with four replicates. The
data were expressed as mean ± standard error. One way
Anova and Dunnet post hoc test was employed for
statistical analysis. Statistical significance was established
at P<0.05 (Yue et al., 2010).
RESULTS
Table 1 shows the wound closure for the all the extracts at
tested concentrations, negative and control groups in fixed
time points. As an early preliminary study only two
concentration of plant extract were used in the treatment.
The diameters of the created wound were calculated using
recommended Axio Vision Release 4.8.1 software.
Among all the extracts, bulb and root produced better cure
for the created wound. The bulb extract 1µg mL-1
produced 100% improvement at 36 h, 92.11% is a control
group, 88.90% is a negative control group and 79.04% 10
µg mL-1 of bulb extract. Even though the wound healed
percentage is nearly similar to bulb extract yet the density
of cell growth is more obvious in bulb compared to
control group. The picture shown clearly the intense
growth of cells compared to control group. Methanolic
root extract displays 100% heals at 36h, while control
group was 79.45%, negative control group 83.74% and 10
µg mL-1 concentration of root extract was 74.80%. Both
these bulb and root extracts significantly repaired the
injured cells within 36h. The anther, stem, flower and
leaves cured the wound only at 48h. Anther and stem
extracts repairs wound 100% at 1µg mL-1 at 48h;
meanwhile flower extract only heals 95.822% and leaves
extract 91.44%. Hence, both bulb and root extracts were
more effective in wound healing activity compared to
other methanolic extracts.
Fig. 1 displayed images of wound healing at 36h for
various H. littoralis extracts at 1 µg mL-1. The images
were corresponding with the table 1 information. figs. 1B
and 1C shown a clear healing property for root and bulb
extracts compared to control (fig. 1A). The other extracts
were shown a slow healing activity for the created wound
compared to root and bulbs. This exhibits the root and

Sreeramanan Subramaniam et al
Pak. J. Pharm. Sci., Vol.31, No.6, November 2018, pp.2537-2543 2539
bulb extracts possess wound healing property compared
to other extracts. fig. 2 shows the wound healing activity
in high concentration 10µg mL-1. Nevertheless at this
concentration the plant extracts fails to express the wound
healing activity. There was no significant closured area
for the created wounds.
DISCUSSION
Wounds have an incredible influence on the healing
healthcare economy (Agyare et al., 2013). In general the
purpose of wound healing management is to heal the
wound as fast as possible with minimal pain and scar
formation (Clark, 1991; Suntar et al., 2010). Wound
healing is a complicated process of cellular and
biochemical interactions involving various cells such as
keratinocytes, fibroblasts and endothelial cells (Tao et al.,
2007; Krishnamoorty 2012). A significant wound healing
is vital for the reestablishment of disrupted functional
status of the skin and disturbed anatomical continuity
(Ramesh and Shanoy, 2013). An agent could treat a
wound in shortest time without scar is always required in
market (Suntar et al., 2010). The H. littoralis showed
significant promotion of wound healing activity in
fibroblast Hs27 cell line treated with root and bulb
methanolic extracts in 36 h.
Table 1: Percentage of wound closure (%) for each extracts at fixed interval time for Hymenocallis littoralis
Percentage of wound closure (%)
Extracts Hour Control Negative control 1 µg mL-1 10 µg mL-1
4 9.11±5.63 13.44±8.62 3.29±1.77 2.23±1.41
8 12.79±9.31 15.08±9.80 6.14±3.24 5.24±4.40
12 21.40±10.54 24.97±13.29 17.84±2.56 11.53±8.01
24 35.81±19.74 33.32±14.95 36.84±10.07 21.69±14.99
36 70.53±11.26 49.21±17.25 83.29±4.88 34.91±14.99
Anther
48 100.00 90.07±19.86 100.00 54.79±9.26
4 6.69±4.28 6.71±2.88 7.38±5.90 6.98±2.40
8 11.60±5.42 15.66±2.99 5.99±3.06 8.30±3.78
12 17.26±8.15 15.26±8.37 13.79±2.31 10.46±4.73
24 45.67±8.57 52.17±13.41 34.71±12.39 12.21± 4.15
36 66.13±7.41 77.45±6.83 74.34±4.71 48.12±4.58
Flower
48 83.46±11.05 94.09±11.81 95.82±8.34 46.58±8 87
4 6.82±7.84 10.56±2.95 7.75±8.47 2.60±2.76
8 12.18±7.16 13.15±4.00 8.17±5.71 5.10±2.04
12 19.28±5.67 38.04±14.99 17.17±10.31 11.62±3.59
24 61.27±11.59 64.86±2.69 69.52±2.77 65.70±2.74
36 92.11±15.79 88.90±13.24 100.00 79.04±4.94
Bulb
48 100.00 100.00 100.00 100.00
4 4.94±6.95 11.40±15.90 4.92±6.91 0.13±4.22
8 12.35±5.82 9.42±4.03 5.57±6.40 6.67±8.68
12 15.94±6.08 24.23±20.04 11.08±7.56 5.88±8.00
24 54.10±11.03 48.26±17.11 24.48±9.98 17.92±6.42
36 70.58±8.38 82.41±11.82 68.95±10.43 41.30±10.40
Leaf
48 89.25±12.73 100.00 91.44±10.84 71.89±5.86
4 5.50±1.95 7.06±7.68 16.06±5.49 7.71±2.53
8 9.00±4.45 11.49±8.00 13.89±4.44 8.04±5.60
12 20.65±6.15 22.95±5.00 23.33±5.41 20.32±5.10
24 56.82±10.17 66.08±9.33 69.00±5.23 56.76±15.65
36 79.45±13.95 83.74±12.53 100.00 74.80 ± 17.41
Root
48 100.00 100.00 100.00 100.00
4 12.37±11.33 7.97±2.74 5.80±2.42 9.35±10.20
8 15.72±11.18 12.06±5.50 10.85±7.91 10.84±9.03
12 25.32±13.29 18.12±3.55 23.56±6.75 16.67±8.25
24 56.11±9.96 59.60±5.49 61.94±8.04 52.55±5.49
Stem
36 78.14±16.26 80.33±13.92 92.89±8.21 74.15±8.27
48 95.10±9.81 100.00 100.00 100.00
Note: Experiment was conducted in quadruplicates (n=4)

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Pak. J. Pharm. Sci., Vol.31, No.6, November 2018, pp.2537-2543
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In vitro scratch-wound assay is an economical and direct
technique to study cell migration in cell lines (Liang et
al., 2007). This assay is very useful since the technique is
very quick to work, can be used to screen wide variety of
samples simultaneously and easily compared to in vivo
wound healing technique (Thakur et al., 2011).
Commonly in in vitro model fibroblast cell growth will be
stimulated and create wound to observe the healing
properties (Adetutu et al., 2011). In tissue repairing
mechanism, the skin fibroblast proliferation is crucial
since it involves migration, proliferation, contractions and
collagen production (Mimura et al., 2004; Adetutu et al.,
2011). Inflammatory cells releases cytokines and growth
factor induce the dermal fibroblast and myofibroblasts in
the wound to initiate the tissue repair mechanism
(Schmidt et al., 2009).
Fig. 1: Wound healing of methanolic extracts Hymenocallis littoralis at 36 hours for 1 µg mL-1. [A] Wounded cells at 0
hour; [B] Bulb extract [C] Root extracts [D] Anther extract [E] Flower extract [F] Leaf extract [G] Stem extract

Sreeramanan Subramaniam et al
Pak. J. Pharm. Sci., Vol.31, No.6, November 2018, pp.2537-2543 2541
The wound healing activity exhibited by H. littoralis is as
per in descending order with the bulb > root > stem >
anther > flower > leaf. The findings were presented in
table 1. fig. 1 displayed images of wound healing at 36 h
for various H. littoralis extracts at 1µg mL-1. The H.
littoralis mean IC50 antitumor activity was 75.72µg mL-1
(Yue et al., 2010) [10]. Thus for wound healing activity 1
and 10µg mL-1 concentration was chosen to be studied.
The root and bulb extracts exhibit wound healing activity.
Saponins, flavonoids, tannins (Obi et al., 2011), alkaloids
and triterpenoids (Udegbunam et al., 2011) are the plant
phytoconstituents attributes for the wound healing
activity. H. littoralis plant extracts possess numerous
isolated alkaloids phytoconstituents from the bulb and
Fig. 2: Wound healing of methanolic extracts Hymenocallis littoralis at 36 hours for 10µg mL-1. [A] Wounded cells at
0 hour; [B] Bulb extract [C] Root extracts [D] Anther extract [E] Flower extract [F] Leaf extract [G] Stem extract

Wound healing activity of Hymenocallis littoralis - Moving beyond ornamental plant
Pak. J. Pharm. Sci., Vol.31, No.6, November 2018, pp.2537-2543
2542
root. Hence, alkaloids constituents in the extracts
influence the wound healing process at tested
concentrations.
Fig. 2 explains the wound healing activity of H. littoralis
extracts images at 36 h for 10µg mL-1. This exhibits that,
higher the concentration of extracts lower curing effects
on the wound. These may be due to high content of
bioactive compounds that lead to reduction of
proliferation activity. Previously, reported presences of
anti-proliferation bioactive substances in H. littoralis
(Thakur et al., 2011). The brine shrimp lethality assay's
results (data not presented here) revealed that the H.
littoralis leaf extract possess a highest cytotoxicity
activity compared to other plant extracts. Thus, this could
explain the reason for slow healing process of the leaf
extract for both 1 and 10µg mL-1 (fig. 1F and 2F).
Therefore, the cytotoxicity effects of the extracts might
influences the wound healing capability.
In the wound healing process the reactive oxygen species
(ROS) are very deleterious due to the harmful effects on
cells and tissues. The ROS degrade the absorbable
synthetic biomaterials in cells (Aliyeva et al., 2004;
Kumar et al., 2007). Cytoprotective enzymes known as
free-radical-scavenging enzymes play a crucial role in
regulating the wound healing mechanism by reducing, de-
activating and removing the ROS from cells (Kumar et
al., 2007). Antioxidant may become therapeutic agents in
wound healing process. The application of antioxidant
compounds is proven to improve significantly the wound
healing activity (Aliyeva et al., 2004; Kumar et al., 2007).
H. littoralis root and bulb show a promising wound
closure percentage at 36 hours of treatment. Bulb has high
phenolic content (28.97mg GAE) and alkaloids may cause
for the favorable wound cures at 36h at 1µg mL-1.
Moreover, this plant has an antibacterial activity against
Gram-positive bacteria Staphylococcus aureus and Gram-
negative bacteria Pseudomonas aeruginosa (Thiem and
Grosslinka, 2003) and demonstrated antifungal activity
against Candida albicans (Sundarasekar et al., 2012) for
this plant. Thus, the Hymenocallis littoralis bulb and root
extracts could be used as wound healing remedy at lower
concentration of 1µg mL-1.
CONCLUSION
This finding demonstrated that Hymenocallis littoralis
bulb and root exhibits a promising wound healing activity
for the scratch assay with Hs27 cell line. These
methanolic extracts displays a closure wound healing
activity at 36h at treated concentration 1µg mL-1.
Methanolic extracts possess phytoconstituents which are
responsible to induce the release of cytokines and growth
factors responsible for the tissue repairing mechanism in
Hs 27 cell line. Further molecular studies currently been
conducted to investigate the involvement of various
mediators such as eicosanoids, prostaglandins, cytokines,
responsible growth factors and transcription factors
protein kinases.
REFERENCES
Abou-Donia AH, Toaima SM, Hammoda HM, Shawky E,
Kinoshita E and Takayama H (2008). Phytochemical
and biological investigation of Hymenocallis litoralis
Salisb. Chem & Biodivers., 5: 332-341.
Adetutu A, Morgon WA and Corcoran O (2011).
Ethnapharmacological survey and in vitro evalaution of
wound-healing plants used in South-western Nigeria. J.
Ethnopharmacol., 137: 50-56.
Agyare C, Dwobeng AS, Agyepong N, Boakye YD,
Mensah KB, Ayande PG and Martin AY (2013).
Antimicrobial, antioxidant and wound healing
properties of Kigelia Africana (Lam.) Beneth. and
Strophantus hispidus DC. Adv. Pharmacol Sci., 2013:
10.
Aliyeva E, Umur S, Zafer E and Acigoz G (2004). The
effect of polylactide membranes on the levels of
reactive oxygen species in periodontal flaps during
wound healing. Biomaterials., 25: 4633-4637.
Clark RAF (1991). Cutaneous wound repairs. In:
Goldsmith LA (ed.) Physiology, Biochemistry and
Molecular Biology of Skin. Oxford University Press,
New York, USA, p.576.
Dalazen P, Molon A, Biavatti MW and Kreuger MRO
(2005). Effects of the topical application of the extract
of Vernonia scorpioides on excisional wounds in mice.
Rev Bras Farmacogn., 15: 82-87.
Fronza M, Heinzmann BM, Hamburger M, Laufer S and
Merfort I (2009). Determination of the wound healing
effect of Calendula extracts using the scratch assay
with 3T3 fibroblasts. J. Ethnopharmacol., 126: 563-
467.
Gurtner GC, Werner S, Barrandon Y and Longaker MT
(2008). Wound repair and regeneration. Nature, 453:
314-321.
Houghton PJ, Hylands PJ, Mensah AY, Hensel A and
Deters AM (2005). In vitro tests and
ethnopharmacological investigations: Wound healing
as an example. J. Ethnopharmacol., 100: 100-107.
Idso SB, Kimball BA, Ill GRP, Garner LC, Pettit GR and
Backhaus RA (2000). Effects of atmospheric CO2
enrichment on the growth and development of
Hymenocallis littoralis (Amaryllidaceae) and the
concentrations of several antineoplastic and antiviral
constituents of its bulbs. J. Bot., 87: 769-773.
Ingrassia L, Lefranc F, Mathieu V, Darro F and Kiss R
(2008). Amaryllidaceae isocarbostyril alkaloids and
their derivatives as promising antitumor agents. Transl.
Oncol., 1: 1-13.
Krishnan P (2006). The scientific study of herbal wound
healing therapies: Current state of play. Curr. Anaesth.
Crit. Care., 17: 21-27.

Sreeramanan Subramaniam et al
Pak. J. Pharm. Sci., Vol.31, No.6, November 2018, pp.2537-2543 2543
Krshnamoorty JR, Sumitira S, Rnjith MS, Gokulshankar
S, Ranganathan S, Mohanty BK and Prabhakaran G
(2012). An in vitro study of wound healing effect of a
poly-herbal formulation as evidenced by enhanced cell
proliferation and cell migration. EDOJ., 8: 1-7.
Kumar B, Vijayakumar M, Govindarajan R and
Pushpangadan P (2007). Ethnopharmacological
approaches to wound healing exploring medicinal
plants of India. J. Ethnopharmacol., 114: 103-113.
Liang CC, Park AY and Guan JL (2007). In vitro scratch
assay: A convenient and inexpensive method for
analysis of cell migration in vitro. Nat Protoc., 2: 329-
334.
Margaret I, Srinivasa RP and Kaiser J (1998).
Antiinflammatory profile of Tridax procumbens in
animal fibroblast cell models. Phytother Res., 12: 285-
287.
Masoko P and Eloff JN (2007). Screening of twenty-four
South African Combretum and six Terminalia species
(Combretaceae) for antioxidant activities. Afr. J. Tradit.
Complement Med. altern., 4: 231-239.
Mimura Y, Ihn H, Jinnin M, Asano Y, Yamane K and
Tamaki K (2004). Epidermal growth factor induces
fibronectin expression in human dermal fibroblast via
protein kinase C delta-signaling pathway. J. Invest
Dermatol., 122: 1390-1398.
Obi RK, Nwanebu FC, Ndubuisi-Nnaji UU, Onuoha LN
and Chiegboka AN (2011). Ethanolic extraction and
phytochemical screening of two Nigerian on pathogens
isolated from wound infections. Pharmacie Globale
(IJCP),10: 1-5.
Ramesh HA and Shenoy DB (2013). Effect of Careya
arborea extracts on wound healing activity in rats.
IJRPBS, 2: 36-43.
Schmidt C, Fronza M, Goettert M, Geller F, Luik S,
Flores, EMM and Merfort I (2009). J.
Ethnopharmacol., 122: 523-532.
Sundarasekar J, Sahgal G and Subramaniam S (2012).
Anti-candida activity Hymenocallis littoralis extracts
for opportunistic oral and genital infection Candida
albicans. Bangladesh J. Pharmacol., 7: 211-216.
Suntar IP, Akkol EK, Yalcin FN, Koca U, Keles H and
Yesilada E (2010). Wound healing potential of
Sambucus ebulus L. and isolation of an active
component, quercetin 3-0-glucoside. J
Ethnopharmacol., 129: 106-114
Tao H, Anthony J, Berno, David RC and Kelly AF (2007).
In vitro human keratinocyte migration rate are
associated with SNPs in the KRT1 interval. PLoS
ONE., 2: e697.
Thakur R, Jain N, Pathak R and Sandhu SS (2011).
Practices in wound healing studies of plants. Evid
Based Complement Alternat Med, Article ID 438056,
17 pages
Thiem B and Grosslinka O (2003). Antimicrobial activity
of Rubus chamaemorus leaves. Fitoterapia., 75: 93-95.
Udegbunam RI, Udegbunam SO and Ugwuanyi SC
(2011). The wound healing potential of the ethanol
extract of Bridelia Ferruginea Benth - In-vivo study. J.
Pharm. Biomed. Sci., 6: 17-21.
Yue PYK, Leung EPY, Mak NK and Wong RNS (2010).
A simplified mehtod for quantifting cell
migration/wound healing in 96-well plates. J. Biomol.
Screen, 15: 427-434.