Therapeutic Effects of Extracts from Spirulina platensis versusBevacizumab on Inflammation-Associated Corneal NeovascularizationAmany Abd El-Fattah El-Shazly1*, Asmaa Ibrahim Ahmed2, Yasser Abdelmageuid Elzankalony1 and Walid Mohamed Abd El Raouf El-Zawahry1

1Department of Ophthalmology, Ain Shams University, Cairo, Egypt2Faculty of Medicine, Department of Anatomy, Ain Shams University, Cairo, Egypt*Corresponding author: Amany Abd El-Fattah El-Shazly, Department of Ophthalmology, Ain Shams University, Cairo, Egypt, Tel: 002 010 1859928; E-mail:amany_elshazly@ymail.com

Received date: Sep 18, 2015; Accepted date: Dec 08, 2015; Published date: Dec 12, 2015

Copyright: © 2015 El-Fattah El-Shazly AA, et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permitsunrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

Abstract

Aim: The aim of this work was to study histological and immunohistochemical changes resulting from cornealalkali burn and to assess these changes after treatment with either polysaccharide extract from Spirulina platensis(PSP) or Bevacizumab.

Methods: Alkali burn was induced in corneas by direct application of a filter paper ring (2.5 mm diameter)saturated with 1 N NaOH and applied to the center of the rat corneas for 30 seconds. PSP (extracted from drypowder of Spirulina platensis) and avastin were administered topically on the corneas 4 times daily for 7 daysstarting 7 days after induction of corneal alkali burn. The therapeutic effects of PSP and avastin were evaluated dailyusing slit-lamp. At the end of the therapy, corneas were harvested for H and E staining, Masson trichrome staining,immunohistochemical study for assessment of inflammation and corneal neovascularization (CorNV).

Results: Topical application of either PSP extract or Avastin had significant therapeutic effects on corneal injuriesinduced by alkali burn. This was demonstrated by decreasing the CorNV, total and differential corneal thickness andinflammatory cellular responses. They increased the epithelial thickness. PSP showed better results than Avastin inthis regards.

Conclusion: The naturally occurring Spirulina platensis (PSP) is more cost effective in treatment of cornealneovascularization and decrease inflammation and fibroblast activity in a rat model of corneal neovascularizationinduced by alkali burn as compared to Avastin.

Keywords: Spirulina platensis; Bevacizumab; Avastin; CornealNeovascularization; Inflammation; Immunohistochemical changes

IntroductionChemical trauma of the eye can range from mild irritation to the

severe damage of ocular surfaces such as the conjunctiva, cornea, andanterior segment. Such injury has the possibility to result in permanentvision loss. Chemical burns of the cornea cause both superficial anddeep neovascularization [1-3] which can lead to significant diminutionof vision due to scar formation and lipid deposition [4]. Regarding theeffect of strong alkalis, ocular tissues have a limited ability to beprotected against such burns, which denature proteins and saponifylipids.

Neovascularization is a poorly understood pathologic response ofthe cornea against chronic inflammation which caused by infection,sterile corneal ulceration, chemical or thermal trauma, or the immunerejection of corneal grafts. Following chemical burns, inflammatorycells such as polymorphonuclear leukocytes, and mesenchymal cellssuch as myofibroblasts, activated keratocytes, macrophages, andneovascularization factors are stimulated. Some of the factorsresponsible for the encouragement of inflammation include vascularendothelial growth factor (VEGF), platelet-activating factor,transforming growth factors, basic fibroblast growth factor, and tumor

necrosis factor-α [5-7]. It has been reported previously that VEGF is upregulated in corneas with inflammation and vascularization and that itis a significant angiogenic factor in corneal neovascularization [8,9].

Bevacizumab (avastin) is a monoclonal antibody that binds toVEGF-A and all its isoforms, its amino acid sequence contains bothhuman IgG (93%) and murine antibody (7%). It has been accepted asan antiangiogenic pharmaceutical for the treatment of certain types ofcancers and it has also been used to treat ocular neovascularization[10]. In numerous studies it has been reported that topical orsubconjunctival Bevacizumab application results in the inhibition orregression of corneal neovascularization [11-13].

Spirulina platensis (PSP) has been proven to possess multiplebioactivities including inhibition of tumor invasion and antiviral,antioxidant, chemoprotective and radioprotective properties [14-16].To our knowledge, this study demonstrates for the first time that PSPhas a strong inhibitory effect on inflammation-induced cornealneovascularization in comparison to Avastin which suggest thepotential use of PSP in the treatment of inflammatoryneovascularization-related corneal diseases.

The aim of this work was to study histological andimmunohistochemical changes resulting from corneal alkali burn andto assess these changes after treatment with either polysaccharideextract from Spirulina platensis (PSP) or Bevacizumab.

El-Fattah El-Shazly AA et al., J Med Surg Pathol

Research Article Open Access

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Material and Methods

Experimental animalsIn this study, 40 adult male albino rats of Sprague Drawly strain

weighing 200-250 g and averaging 16 weeks old were utilized. Theywere obtained from the Animal house Ain shams University faculty ofmedicine. Animals were maintained according to the principle andguidelines of the Committee for the Purpose of Control andSupervision of Experiments on Animals (CPCSEA).

Food and tap water were available ad libitum. In the windowlessanimal area automatic temperature (21 ± 1°C) and lighting controls(12 h light/12 h dark cycle) were done. Humidity ranged from 55% to60%. All animals received human care according to the principlesdefined in the “Guide for the Care and Use of Laboratory Animals”prepared by the National Academy of Sciences and published by theNational Institutes of Health.

Corneal alkaline burn injuryUnder general anesthesia with intraperitoneal Xylazine

hydrochloride (5 mg/kg) (Rompun®, Bayer, Turkey) and ketaminehydrochloride (50 mg/kg) (Ketalar®, Eczacibasi, Turkey), the alkaliburn CorNV model was established by instillation of topical anesthesiawith a drop of 0.5% proparacaine hydrochloride (Alcaine eye drops;Alcon Inc., Fort Worth, TX) and direct application of a filter papercircle (2.5 mm diameter) was soaked with 1 N NaOH and applied tothe corneal center for 30 s. The cornea was then rinsed with saline (60mL) for 30 s, according to the method created by Ormerod andcolleagues with modification [17]. The burn stimulus response wasscored by slitlamp inspection, as grade 0 (no blister), +1 (small blister),+2 (medium blister), or +3 (large blister), according to lesions detectedon the corneal surface [18].

Following cauterization, the burn stimulus scores were immediatelycalculated and treatment was begun with gentamicin eye drops fourtimes a day for one week.

Experimental designThe animals were classified into four groups, ten rats each, as the

following:

Group 1 (control group): 10 rats, were divided in to 2 sub-groups

Sub group 1A: 5 rats (10 eyes) to whom balanced saline solution(BSS) was administered topically on the corneas 4 times daily for 14days

Sub group 1B: to assess drug toxicity without alkali burn: 5 rats (10eyes) will be subdivided as the following:

• Their (Right) Rt eye received Bevacizumab (Avastin) topically 4times daily for 7 days

• Their (Left) Lt eye received PSP extract topically on the corneas 4times daily for 7 days

Group 2 (corneal alkali burned group): 10 rats (20 eyes) to whomthe alkali burn CorNVs model was established.

Group 3 (Bevacizumab [Avastin]-treated group): 20 rats (40 eyes),were divided in to 2 sub-groups:

Sub group 3A: Their Rt eye received Bevacizumab (Avastin)topically 4 times daily starting 7 days after induction of corneal alkaliburn for 7 days

Sub group 3B: Their Lt eye received PSP extract was administeredtopically 4 times daily starting 7 days after induction of corneal alkaliburn for 7 days

All experimental animals were carried out following the guidelinesof the Association for Research in Vision and OphthalmologyStatement for the Use of Animals in Ophthalmic and Vision Research.

Drug preparation and treatment protocolPreparation of polysaccharide extract from Spirulina platensis

(PSP): PSP was prepared as described previously [19]. In brief, drypowder of was incubated with 95% (V/V) ethanol overnight, torrefied,and re-incubated in NaOH (pH 10.0) solution at 80 for 4-6 hours, thencentrifuged and collected the supernatant, adjusted pH to 7.0,precipitated with 5% trichloracetic acid (TCA) at 4 overnight,centrifuged and collected the supernatant, then precipitated with 5%TCA for another 3 hours, centrifuged and the supernatant wasprecipitated with ethanol (1/5:V/V) at 4 overnight. The precipitate waswashed twice with acetone and then was lyophilized in a freeze dryer,and stored at -20°C. This precipitate, mainly containingpolysaccharides (PSP), was dissolved in normal saline and filtered with0.22 µm filtration membrane and the stock polysaccharidesconcentration was measured using anthrone-sulfuric acid method. Fortopical application, the stock PSP was adjusted to a concentration of 2mg/ml in sodium chloride eye drops compound.

Preparation of Bevacizumab eye drops: Bevacizumab eye dropswere prepared in the hospital pharmacy by dilution with normal saline(0.9%) under sterile situations to a concentration of 10 mg/ml andwere kept at 4°C. The eye drops were stored at -20°C for up to 14 days;after opening, they were used within 1 day and kept at 4°C during thattime. All procedures were performed by the same investigator.

Clinical monitoring: Assessment of corneal inflammation andneovascularization: A slit-lamp microscopy examination wasperformed under anesthesia and analgesia, on 1st day to assess theburn stimulus response, on the 7th day to evaluate the cornealneovascularization and on the 14th days to evaluate the therapeuticeffects of PSP extract and Bevacizumab. Then we evaluatedinflammation and neovascularization by image analysis as it is anobjective tool using the stained sections [20,21].

Histological analysisThe rats were sacrificed on the 15th day using a high dose of

Pentothal sodium (Pentothal®, Abbott, Italy). The globes were thenenucleated and fixed in 10% buffered formalin for 24 h. Corneas werethen removed from the limbus and 5 μm thick paraffin sections wereset and stained with hematoxylin and eosin (Figure 1) and maissontrichrome (MTC) (Figure 2), then were examined using lightmicroscopy. Sections were evaluated by image analysis to demonstratethe corneal thickness, the amount of neovascularization, the quantityof inflammation, and the fibroblast activity. Image analysis wasperformed by an examiner who does not know the coding of the studygroups to avoid bias.

Citation: El-Fattah El-Shazly AA,Ibrahim Ahmed A, Elzankalony YA,Abd El Raouf El-Zawahry WM (2016) Therapeutic Effects of Extracts fromSpirulina platensis versus Bevacizumab on Inflammation-Associated Corneal Neovascularization. J Med Surg Pathol 1: 102.

doi:

10.4172/jmsp.1000102

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Volume 1 • Issue 1 • 1000102J Med Surg Pathol, an open access journalISSN:2472-4971

10.4172/2472-4971.1000102doi:

ImmunohistochemistryAssessment of angiogenesis by immunohistochemistry of vascular

endothelial growth factor (VEGF) and assessment of fibroblast activityusing α-smooth muscle actin.

As paraffin sections were stained with avidin-biotin peroxidase fordemonstration of cells immunoreactive to VEGF and α-smooth muscleactin and counterstained with HX (Figures 4 and 5). Antibodiesagainst VEGF and α-smooth muscle actin were purchased from (SantaCruz Biotechnology, USA).

Image analysisSections were examined by using Leica DM2500 microscope with

built in camera (Weltzelar, Germany). All images were digitallyacquired using an image analyzer Leica Q win V.3 program (Weltzelar,Germany) installed on a computer in histology department faculty ofmedicine Ain Shams University.

Ten different non-overlapping fields from ten different stainedsections of ten different rats were examined in each group formeasuring each of the following:

• Corneal epithelial layer thickness• Substantia propria (stroma) layer thickness• Descemet's membrane (DM) layer thickness• The total corneal thickness• Mean Mononuclear cells infiltration count• Active fibroblast cell infiltration count• Corneal neovascularization (area% of brown color) using VEGF

All measurements were taken at high power field of magnification(400X), all data are collected, revised and subjected for StatisticalAnalysis.

Statistical analysisStatistical analysis was done using (Statistica software, version 8).

Quantitative variables were expressed as mean ± SD and qualitativevariables were given as numbers and percentage. Student’s t-test wasused to assess the statistical differences regarding the quantitativevariables whereas ANOVA test was used for the qualitative ones.

ResultsBy slit-lamp microscopy examination revealed that the mean burn

stimulus score was (2.25 ± 0.79 µm) for group 2, (2.10 ± 0.64 µm) forgroup 3A and (2.15 ± 0.59 µm) for group 3B. No statistical differenceswere found between the different groups (p=0.52).

Group 1 B show no deleterious effect was seen on PSA on thecornea, exactly similar to avastin results.

Histological resultsIn hematoxylin and eosin stained sections, the superficial layer of

stroma show aggregation of polymorphonuclear cell infiltrationsnoticed in group 2 with new vessel formation. These changes were fewin group 3A and absent in group 3B (Figure 1). In Masson's trichrome(MTC) sections, abnormal disorganized loose collagen fibers presentin corneal stroma with new vessels in group 2, this disorganization wasless observed in group 3A while group 3B apparently similar to the

control group, also there is difference in the corneal thickness betweenthe different studied groups (Figure 2). Immunohistochemical stainedsections for assessment of neovascularization using VEVF showedpositive immune reaction in group 2, group 3A presented weakreactions while group 3B showed negative reaction (Figure 4). α-smooth muscle actin stained sections showed positive brown stainedfibroblasts in group 2, weak staining in group 3A and negative reactionin group 3B (Figure 5).

Figure 1: Dark stained nuclei of mononuclear cell infiltrate in thesubepithelial layer (red arrow) and CorNV (*) is present in group 2while group 3A show few vessel and absent in group 3B (HandE X400).

Figure 2: Total corneal thickness in histological Masson's trichromestaining sections in the different four studied groups. Notice loosedisorganized collagen fibers (red arrow) present in corneal stromawith CorNV (*) in group 2, this disorganization was less observedin group 3A while group 3B apparently similar to the control group.(400X).

Citation: El-Fattah El-Shazly AA,Ibrahim Ahmed A, Elzankalony YA,Abd El Raouf El-Zawahry WM (2016) Therapeutic Effects of Extracts fromSpirulina platensis versus Bevacizumab on Inflammation-Associated Corneal Neovascularization. J Med Surg Pathol 1: 102.

doi:

10.4172/jmsp.1000102

Page 3 of 7

Volume 1 • Issue 1 • 1000102J Med Surg Pathol, an open access journalISSN:2472-4971

10.4172/2472-4971.1000102doi:

Group 1 Group 2 Group 3A Group 3B p value

1 vs 2 1 vs 3A 1 vs 3B 2 vs 3A 2 vs 3B 3A vs 3B

Epithelialthickness (µm)

20.01

± 2.23

14.25

± 2.01

21.65

± 0.88

20.78

± 1.18

<0.001 <0.001 0.16 <0.001 <0.001 0.02

Stromalthickness (µm)

98.47

± 2.90

192.91

± 9.97

115.52

± 2.53

104.34 ±2.04

<0.001 <0.001 0.02 <0.001 <0.001 <0.001

DM thickness(µm)

2.65

± 0.28

3.07

± 0.28

2.81

± 0.27

2.64

± 0.26

<0.001 0.09 0.91 <0.001 <0.001 0.04

Total cornealthickness (µm)

122.15

± 4.95

211.04

± 4.38

124.42

± 2.71

119.32 ±2.82

<0.001 0.10 0.11 <0.001 <0.001 0.001

Table 1: Different corneal layer thickness (µm) in the different four studied groups.

As Fuentes-Julián et al. (2015) stated that one of the signs ofinflammation is corneal edema with increased thickness, We studiedthe different corneal layer thickness in histological H and E (Figures 1and 2) staining sections by Image analysis and we found that [22]:

Epithelial thickness in group 1 was statistically higher than in group2 (p ≤ 0.001), and lower than in group 3A (p ≤ 0.001). However it wasnot statistically different from group 3B (p=0.16). Also group 3A wasstatistically higher than in group 2 (p ≤ 0.001) and group 3B wasstatistically higher than in group 2 (p ≤ 0.001). Also group 3A wasstatistically higher than in group 3B (p=0.02) (Table 1)

Stromal thickness in group 1 was statistically lower than in group 2(p ≤ 0.001), group 3A (p ≤ 0.001) and group 3B (p=0.02). Also group 2was statistically lower than in group 3A (p≤0.001) and group 2 wasstatistically lower than in group 3B (p ≤ 0.001). Also group 3A wasstatistically higher than in group 3B (p ≤ 0.001) (Table 1)

DM thickness in group 1 was statistically lower than in group 2 (p ≤0.001), but there is no statistical different between group 1 and group3A (p=0.09) and group 3B (p=0.91). Also group 2 was statisticallyhigher than in group 3A (p ≤ 0.001) and group 2 was statisticallyhigher than in group 3B (p ≤ 0.001). Also group 3A was statisticallyhigher than in group 3B (p=0.04) (Table 1)

Finally, the total corneal thickness in group 1 was statistically lowerthan in group 2 (p ≤ 0.001), but there is no statistical different betweengroup 1 and group 3A (p=0.10) and group 3B (p=0.11). Also group 2was statistically higher than in group 3A (p ≤ 0.001) and group 2 wasstatistically higher than in group 3B (p ≤ 0.001). Also group 3 wasstatistically higher than in group 3B (p=0.001) (Table 1).

Corneas of eyes with alkali burn, possessed more new vessels, whilethat treated with Bevacizumab group, and that treated Spirulinaplatensis (PSP) group with statistical significant difference betweengroup 2 and group 3A (p ≤ 0.001) and between group 2 and group 3B(p ≤ 0.001). Also there was statistical significant difference betweengroup 3A and group 3B (p=0.02) (Figures 5 and 6) (Table 2). The levelsof vascular endothelial cells were examined by immunohistochemistryfor vascular endothelial growth factor. The results showed that vascularendothelial growth factor were prominent in the burned corneal

stromas of control rat whereas that of Avastin and PSP-treated corneasexhibited negligible staining (Figures 4 and 5).

Figure 3: Comparison between the Bevacizumab (Avastin)-treatedgroup (group3A) and Spirulina platensis (PSP)–treated, group(group 3B) regarding the total corneal thickness.

Also histological H and E staining showed that the corneas of eyeswith alkali burn, showed more mononuclear cells infiltration in thecorneal stroma than did corneas treated with Bevacizumab and PSP ,with statistical significant between group 2 and group 3A (p=0.03),group 2 and group 3B (p ≤ 0.001) and group 3A and group 3B (p=0.01)(Table 2).

Histological H and E staining showed that the corneas of group 2,showed more active polymorphonuclear cell infiltration in the cornealstroma than did corneas treated with Bevacizumab and PSP, withstatistical significant difference between group 2 and group 3A (p ≤0.001 ), group 2 and group 3B (p ≤ 0.001). But there is no statisticalsignificant difference between group 3A and 3B (p=0.4) (Table 2).

Group 1 Group 2 Group 3A Group 3B p value

Citation: El-Fattah El-Shazly AA,Ibrahim Ahmed A, Elzankalony YA,Abd El Raouf El-Zawahry WM (2016) Therapeutic Effects of Extracts fromSpirulina platensis versus Bevacizumab on Inflammation-Associated Corneal Neovascularization. J Med Surg Pathol 1: 102.

doi:

10.4172/jmsp.1000102

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1 vs 2 1 vs 3A 1 vs 3B 2 vs 3A 2 vs 4 3A vs 3B

Neovascularization (percentage/high power field)

0.20 ± 0.11 3.62 ± 0.52 1.26± 0.16 0.99 ± 0.42 <0.001 <0.001 <0.001 <0.001 <0.001 0.02

Mononuclear cellsinfiltration (perhigh power field[Magnification,×400.])

0.00 ± 0.00 3.85 ± 1.31 2.25 ± 2.55 0.50 ± 0.51 <0.001 <0.001 <0.001 0.04 <0.001 0.01

Active fibroblastscell infiltration (perhigh power field[Magnification,×400.])

5.90 ± 3.63 22.00 ± 2.81 6.50 ± 2.12 5.60 ± 3.50 0.00000 0.5 0.3 <0.001 <0.001 0.4

Table 2: Corneal neovascularization and different inflammatory cell infiltration in the different four studied groups.

Figure 4: VEGF were prominent in the burned corneal stromas of control rat whereas that of Avastin and PSP-treated corneas exhibitedinsignificant staining. Also there is positive immune reaction in group 2, group 3A presented weak reaction while group 3B showed negativereaction.

Discussion

Baseline assessmentThe mean burn stimulus score was the same between the studied

groups has no statistical difference (p=0.48) and (p=0.67). This makesit plausible to compare between these groups after therapy.

Post therapy assessmentCorNVs, as assessed immunohistochemically for vascular

endothelial growth factor, was significantly lower in treated groupscompared to untreated one, as CorNV is a significant sight-threateningevent that usually leads to loss of central vision or even results inblindness [19].

Inflammatory reactions were assessed through assessment ofthickness of different corneal layers shown in histological Masson'strichrome staining sections as well as quantification of inflammatory

cells shown in H and E and α smooth muscle acting stained stainingsections [22].

Total corneal thickness was significantly lower in Bevacizumab(Avastin) treated group and Spirulina platensis [PSP] treated groupcompared to untreated alkaline burn group.

Similarly, epithelial thickness was significantly higher inBevacizumab (Avastin) treated group and Spirulina platensis [PSP]treated group) compared to untreated alkaline burn group.

Moreover, stromal thickness was significantly lower in Bevacizumab(Avastin) treated group and Spirulina platensis [PSP] treated groupcompared to untreated alkaline burn group.

DM thickness was significantly lower in Bevacizumab (Avastin)treated group and Spirulina platensis [PSP] treated group (comparedto untreated alkaline burn group.

Citation: El-Fattah El-Shazly AA,Ibrahim Ahmed A, Elzankalony YA,Abd El Raouf El-Zawahry WM (2016) Therapeutic Effects of Extracts fromSpirulina platensis versus Bevacizumab on Inflammation-Associated Corneal Neovascularization. J Med Surg Pathol 1: 102.

doi:

10.4172/jmsp.1000102

Page 5 of 7

Volume 1 • Issue 1 • 1000102J Med Surg Pathol, an open access journalISSN:2472-4971

10.4172/2472-4971.1000102doi:

Figure 5: α smooth muscle actin stained sections showed positivebrown stained fibroblasts in group 2, weak stain in group 3A andnegative stain in group.

Figure 6: Comparison between the Bevacizumab (Avastin)-treatedgroup (group 3A) and Spirulina platensis (PSP)-treated, group(group 3B) regarding corneal neovascularization represented inpercentage.

Fibroblasts cellular infiltration was found to be significantly lower inBevacizumab (Avastin) treated group and Spirulina platensis [PSP]treated group compared to untreated alkaline burn group.

Mononuclear cells infiltration was found to be significantly lower inBevacizumab (Avastin) treated group and Spirulina platensis [PSP]treated group compared to untreated alkaline burn group.

The previous results demonstrate the beneficial outcome ofBevacizumab (Avastin) and Spirulina platensis [PSP] on inflammatoryprocess and regression of CorNVs induced by alkali burn.

In fact, Bevacizumab (avastin) effect was previously studied byOzdemir et al. (2014), who demonstrated that topical administrationof Bevacizumab inhibits corneal neovascularization and decreasesinflammation and fibroblast activity in a rat model of cornealneovascularization induced by alkali burn [23]. Öner et al. (2012),stated that topical use of Bevacizumab is effective and safe incontrolling corneal neovascularization [24]. Hashemian et al. (2011),found that topical Bevacizumab can prevent CorNV in rats [25]. AlsoKim et al. (2013), proved that topically administered Bevacizumab hasstanding anti-angiogenic effect on corneal neovascularizationfollowing chemical injury in rats [26].

On the other hand, Spirulina platensis [PSP] was studied by Yang etal. (2009), who found that polysaccharide extract from Spirulinaplatensis is a potent inhibitor of CorNVs and that it may be of value inthe therapy of corneal diseases including neovascularization andinflammation. Similarly, Yang et al. (2012), stated that polysaccharideextract from Spirulina platensis [PSP] inhibited alkali burn-inducedinflammation and CorNV more effectively than AME extract at thestudied doses, thus may be used for the treatment of corneal diseasesinvolving neovascularization and inflammation.

Chen et al. (2004), demonstrated the possible anti-inflammatoryeffects of PSP on alkali-burned corneas. The inflammation factors asSDF1 and TNF-α are known to be one of the key regulators ofinflammation and can mediate angiogenesis. In corneas, theinfiltration of SDF1-positive inflammatory cells and the expression ofSDF1 and TNF-α RNA were significantly depressed by PSP [27].

Yang et al. (2009), stated that PSP suppressed the expression ofVEGF, matrix metalloproteinase (MMP2, and MMP9) and stimulatedthe expression of PEDF, suggesting that PSP may inhibit CorNVs bydown regulating the expression of angiogenic factors and up regulatingthe expression of the anti-angiogenic factors. The anti-angiogeniceffects of PSP were mediated by interference with the proliferation,migration, and tube formation of vascular endothelial cells in vitro.

Adisakwattana et al. (2013), reported that hydroxyl cinnamic acidand their derivatives of PSP extract induce corneal epithelial woundhealing by inhibiting protein tyrosine phosphatases (PTP)-1β gene[28]. Also Bodet et al. (2008), stated that narigenin is an effectiveinhibitor of the pro-inflammatory cytokine (IL-1β, IL-6, IL-8 andTNF-α) response stimulated by lipopolysaccharide in bothmacrophages and in whole blood [29].

Comparison of the two treated groups demonstrated superiority(not only non-inferiority basis) of Spirulina platensis [PSP] overBevacizumab [avastin] therapy. This is shown by comparison betweenBevacizumab [avastin] treated group and Spirulina platensis [PSP]treated group. This showed significantly lower stromal thickness, DMthickness, total corneal thickness, CorNVs and active fibroblasts andmononuclear cell infiltration.

Another point in favor of Spirulina platensis [PSP] over Avastin wasits potential to bring down the stromal thickness near to normalcontrol values more than Avastin.

Citation: El-Fattah El-Shazly AA,Ibrahim Ahmed A, Elzankalony YA,Abd El Raouf El-Zawahry WM (2016) Therapeutic Effects of Extracts fromSpirulina platensis versus Bevacizumab on Inflammation-Associated Corneal Neovascularization. J Med Surg Pathol 1: 102.

doi:

10.4172/jmsp.1000102

Page 6 of 7

Volume 1 • Issue 1 • 1000102J Med Surg Pathol, an open access journalISSN:2472-4971

10.4172/2472-4971.1000102doi:

ConclusionThe naturally occurring Spirulina platensis (PSP) is more cost

effective in treatment of corneal neovascularization and decreaseinflammation and fibroblast activity in a rat model of cornealneovascularization induced by alkali burn as compared to Avastin.

ContributorsAmany Abdel-Fattah El-Shazly: data collection, statistics,

manuscript writing and review, Clinical monitoring and surgicalmaneuver. Asmaa Ibrahim Ahmed: examination of histologicalsections and interpretation of data and manuscript review; YasserAbdelmageuid Elzankalony: manuscript review and data collection.Walid Mohamed Abd El Raouf El-Zawahry: data collection,manuscript review.

FundingNo funding. Costs were the responsibility of the authors and

instruments used in the study belong to Faculty of Medicine, a part ofAin Shams University, which is a public governmental organization.

Competing interestsThere is no conflict of interest of any of the authors with any

establishment having a relation to this present work.

Ethics approvalInstitutional Review Board.

Data sharing statementAll data collected here are contained within the manuscript.

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Citation: El-Fattah El-Shazly AA,Ibrahim Ahmed A, Elzankalony YA,Abd El Raouf El-Zawahry WM (2016) Therapeutic Effects of Extracts fromSpirulina platensis versus Bevacizumab on Inflammation-Associated Corneal Neovascularization. J Med Surg Pathol 1: 102.

doi:

10.4172/jmsp.1000102

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Volume 1 • Issue 1 • 1000102J Med Surg Pathol, an open access journalISSN:2472-4971

doi:10.4172/2472-4971.1000102