Amnion Group Research Articles

Comparison of freeze dried human amniotic membrane and its combination with platelet rich fibrin for the epithelial acceleration in an acute wound excision using Rattus Novergicus study model

Link of Video Abstract: https://youtu.be/FvU45HL3HyE Introduction: Human amniotic membrane and platelet rich fibrin (PRF) are biological materials that can accelerate wound healing through their content of growth factors and cytokines. This study aimed to determine the effectiveness of the combination of these two biological agents for wound healing. Methods: This study used an experimental method with a randomized post-test only control group design, to compare the use of human amniotic membrane alone with a combination of human amniotic membrane and PRF in acute full thickness wounds of the skin of Wistar rats. The research subjects were 32 rats which were divided into four groups. Groups 1 and 2 used human amniotic membrane, while groups 3 and 4 used a combination of human amniotic membrane and PRF, then the epithelialization process was observed on the 3rd and 6th day. Result: There was no statistically significant difference in the speed of epithelialization of excision wounds on the backs of rats on the third day (Significance 0.498) and Sixth (Significance 0.9) between the Amnion group and the combination of amnion and PRF because it was greater than α = 0.05. Conclusion: The use of a combination of freeze dried human amniotic membrane and platelet rich fibrin was not significantly more effective than freeze dried human amniotic membrane in accelerating the epithelialization of acute full thickness excision wounds in rats (Rattus novergicus).

Comparison of the outcome of amnion versus conventional (Vaseline-impregnated gauze) dressing in superficial partial thickness burn patients in terms of healing time and mean pain.

Objectives: To compare the healing time and mean pain score of Amnion versus Conventional (Vaseline-impregnated gauze) dressing in superficial partial thickness burn patients. Study Design: Randomized controlled study. Settings: Plastic & Reconstructive Surgery Department, Jinnah Hospital & Burn Center, Lahore. Period: January 2018 to January 2019. Material & Methods: A total of 60 patients with superficial second degree burns full filling the inclusion criteria were recruited for the study. Subjects were divided into two groups randomly. All patients were followed up regularly and dressing was changed on alternate days in Group B (Vaseline-impregnated gauze) and only secondary dressing was changed in Group A (amnion) until the auto sloughage (self-removal) of amnion. Outcome variables i.e. pain during first dressing change and healing time were noted. Statistical analysis of data was done using SPSS version 22. Quantitative variables i.e. age, duration of burn, total body surface area, pain during dressing and healing time were presented as mean and standard deviation. Frequency and percentage was calculated for qualitative variables like gender. The pain during dressing change and healing time of both groups were compared for difference. Student’t’ test was applied to compare the outcome and p-value ≤0.05 was considered as significant. Results: Out of sixty cases, 18 (30.0%) were females and 42 (70.0%) were males, with female to male ratio of 1:2.3. Mean age of patients in group A was 33.0 ± 10.19 years and in group B was 33.73 ± 9.55 years. The mean pain score in group A (amnion group) was 1.93 ± 0.91 and in group B (Vaseline-impregnated gauze) was 3.33 ± 1.56 with p-value of 0.0001. The mean healing time in group A (amnion group) was 15.73 ± 2.79 days and in group B (Vaseline-impregnated gauze) was 22.80 ± 4.44 days with p-value of 0.0001. Conclusion: Amnion dressing in superficial partial thickness burn patients is more effective in terms of mean pain score and healing time as compared to conventional (Vaseline-impregnated gauze) dressing.

Hierarchical cluster analysis in the study of the effect of cytokine expression patterns on endometrial repair and receptivity after hysteroscopic adhesiolysis.

BackgroundIn a previous study, we reported that amnion promotes endometrial cell growth by regulating cytokines. In this study, hierarchical cluster analysis enabled the evaluation of cytokine expression changes after amnion treatment to be explored by cluster patterns. The role of IL1B on endometrial repair and receptivity was revealed.MethodsA total of 30 patients were recruited in this clinical trial (NCT02496052) of hysteroscopic adhesiolysis. They were randomly allocated into an amnion grafts group (amnion group) and a control group. After hysteroscopic adhesiolysis, a Foley catheter covered with a sterilized freeze-dried amnion graft was inserted into the uterine cavity of the participants in the amnion group, whereas for the control group, a Foley catheter without amnion graft was inserted. After surgery, patient follow-up was done for a year. Uterine exudates were collected every day for seven days after surgery, and analyzed by enzyme-linked immunosorbent assays. Hierarchical cluster analysis was performed to compare expression patterns of each cytokine. Single-gene gene set enrichment analysis and differentially expressed genes enrichment analysis of IL1B were performed using NCBI GEO (N=151) to evaluate its potential mechanisms and impact on endometrial receptivity.ResultsCompared to the control group, cytokine expression patterns of the amnion group revealed significant stratifications, which were highly correlated with the expression levels of IL1B on the sixth to seventh day after surgery, improving the pregnant rate. Wilcoxon test revealed significantly low expression levels of IL1B in the reduced endometrial receptivity group compared to the normal group. Moreover, gene set enrichment analysis showed that lysosomes, cell cycle, and calcium signaling pathways were associated with the biological processes in which IL1B plays a role. Screening and enrichment analyses of differentially expressed genes further verified the mechanisms of action of IL1B on endometrial repair and receptivity recovery.ConclusionsAmnion promotes endometrial repair and receptivity by altering the expression levels and patterns of IL1B. Furthermore, by affecting lysosomal, cell cycle, and calcium signaling pathways, IL1B may be one of the factors involved in endometrial repair and receptivity recovery.

Echogenic particles in the amniotic fluid of term low-risk pregnant women: does it have a clinical significance?

In this study, we aimed to analyse the clinical features of the third-trimester pregnant women, with echogenic amniotic fluid and to compare their obstetric and neonatal outcomes with pregnant women with normal amniotic fluid echogenicity. This case-control study was conducted in a tertiary antenatal care centre. A total of 560 term (37–42 weeks of gestation) singleton women; 280 with echogenic particles in amniotic fluid and 280 with clear amniotic fluid, who delivered within 24 h after the ultrasound scan were evaluated. The women in the two groups were similar in terms of age, parity, body mass index, foetal birth weight, and gestational age. More patients in the particulate amnion group had lower Apgar scores (<7) in 1st and 5th minutes than controls (p = .006, p = .031 respectively) however the rate of admission to neonatal intensive care was similar. Vernix stained amniotic fluid was more common in the study group (48.8%, p = .031), the rate of meconium-stained amniotic fluid was similar in the study and control groups (9.6–9.2%, p = .881). The primary caesarean section rate was higher in women with particulate amnion (18.4%, p = .037). Echogenic particles in the amniotic fluid in the third trimester could not be attributed to meconium, however, higher rates of primary caesarean section may require further attention. IMPACT STATEMENT What is already known on this subject? Previous studies showed that high-density intra-amniotic particles were possibly related to vernix caseosa, intra-amniotic bleeding, and meconium. The number of study groups in these studies was also limited. What do the results of this study add? Additional to other previous studies, we found an increased rate of intra-amniotic echogenic particles in male foetuses. What are the implications of these findings for clinical practice and/or further research? The presence of echogenic particles on ultrasound was not related to increased risk for the presence of meconium. Significantly more neonates born to mothers with intra-amniotic echogenic particles tended to have lower Apgar scores (<7), however, this significant difference did not affect the need for NICU admission. The presence of echogenic particles in the amniotic fluid of the third-trimester pregnant women could not be attributed to meconium and adverse perinatal outcomes, however, the higher rates of primary caesarean section may require further attention.

Comparing the results of duraplasty using amniotic membrane versus pericranium as dural graft; concerning CSF leakage and pseudomeningocele

Purpose: In many brain surgeries, dura cannot be primarily repaired and it is necessary to use a graft. Appropriate repair of dura plays a major role in preventing CSF leakage and meningitis. In the cases where sufficient Pericranium is not available for recovery, we need to use graft from other resources. The present study compares the results of amniotic membrane dural graft and pericranium dural graft in terms of CSF leakage and Pseudomeningocele. Materials and methods: This is a semi-empirical research. Thirty cases underwent dural graft with amnion membrane because of inadequate local pericranium for repair. There were 30 other historical cases that had undergone auto-graft dural graft with Pericranium in the same center.Results: The average age of the participants in Pericranium graft group (P, n = 30) and those in amniotic graft group (A, n = 30) was 39.1 ± 15 and 43.9 ± 19 years old, respectively. As for the P and A groups, there were 14 and 17 males, respectively. Two cases of CSF leakage (7%) were observed in P group, while none was reported in A group. There were 3 cases (10%) of meningitis in Pericranium group and 2 cases (7%) were reported in amnion group. Out of 2 cases of meningitis in group P, 2 cases were associated with CSF leak. 5 cases of Hydrocephalus (17%) were observed in group P and 9 cases (30%) were reported in group A. No significant difference was observed between the two groups in any complication. The frequency of hydrocephalus was significantly higher in posterior fossa craniotomy.Conclusion: Amniotic membrane may provide a good source for cases with a large dural defect and can be utilized for all ages. Of course, its application in cases of posterior fossa craniotomy needs to be carefully studied so that the best results may be obtained.

Amniotic Allograft Implantation for Midface Aging Correction: A Retrospective Comparative Study with Platelet-Rich Plasma

Amniotic allografts are becoming more popular for use in soft tissue growth in many areas of medicine because of their immunoprivilege that allows them to proliferate into tissues without rejection by the host. Platelet-rich plasma (PRP) has crossed over from wide orthopedic uses to the aesthetic market for hair restoration and midface volume replacement, owing, in part, to the minimal risk associated with the procedure and the convenience of in-office application. In addition, growth factors provided by PRP help stimulate collagen synthesis in the aging face. However, the potential recruitment of the patient’s own mesenchymal stem cells to the PRP injection site would produce the most favorable and sustained aesthetic outcome. With the advancement of amniotic allograft procedures, the introduction of live mesenchymal cells of the amniotic membrane into the aging midface could be performed in-office similarly to the PRP treatment. This retrospective chart review compares aspects of the amniotic allograft procedure (office time, level of comfort, and downtime) with the aesthetic results of injection into the midface of those undergoing PRP therapy. Analysis of the changes to midface volume, specifically the Ogee curve, observed in the chronological progression of photographs illustrates aesthetic improvements in both PRP and amnion allograft treatment groups, with changes in the facial grading scale. Less patient downtime and slightly more rapid improvements were noted in the amnion group in comparison with the PRP treatment participants.Level of Evidence IV This journal requires that authors assign a level of evidence to each article. For a full description of these Evidence-Based Medicine ratings, please refer to the Table of Contents or the online Instructions to Authors www.springer.com/00266.

The study on the safety and efficacy of amnion graft for preventing the recurrence of moderate to severe intrauterine adhesions

Transcervical resection of adhesion (TCRA) is the standard treatment for the intrauterine adhesions, but the recurrence of adhesions is a tough problem for the gynecologist. In addition, the therapeutic strategy after TCRA about prevention of recurrence remains controversial especially for the patients with moderate to severe intrauterine adhesions (IUAs). Hence, we designed this study to explore the safety and efficacy of fresh amnion grafts for preventing the recurrence after TCRA for patients with moderate to severe IUAs. One hundred patients with moderate to severe IUAs who presented with a history of hypomenorrhea, amenorrhea and infertility were included in the study from January 2015 to December 2017. Patients were divided into amnion group (52 patients) and chitosan group (48 patients). Fresh amnion grafts or intrauterine injections of chitosan were administered after TCRA. Transvaginal ultrasonography (TVUS) and hysteroscopy were performed at the first and third month after the operation. The surgical procedures for all patients were completed successfully without relevant complications. In amnion group, 8 patients exhibited relapse in the first month and 2 patients in three months after surgery; in chitosan group, 23 women exhibited relapse in the first month and 18 patients in three months after surgery. Statistical analysis revealed that the recurrence rate of adhesion in amnion group was significantly lower than those of chitosan group in the first and three months after surgery (P1 = 0.000, P2 = 0.000). After TCRA, fresh amnion graft plays a significant role in preventing further adhesions than injections of chitosan.

Histological Comparison of New Bone Formation Using Amnion Membrane Graft Versus Resorbable Collagen Membrane: An Animal Study.

The purpose of this article was to evaluate the bone induction effects of an amnion membrane-protected graft compared with a collagen membrane-protected graft in the repair of tibial bony defects in dogs. This study was performed using the tibial bone of dogs. After the removal of periosteum, similar holes were made with a 16-mm trephine drill (38 holes in total). For the study group, 10 holes were covered by absorbable collagen and 16 holes by amniotic membrane. In the control group, 12 holes were made and covered by the overlying soft tissue. Tibial bones were exposed after 6 and 12 weeks, and the samples were harvested and histologically processed. New bone formation was evaluated by histomorphometric study. Four Iranian mixed dogs older than 1.5 years were included in this study. The new bone formation was less in the control group when compared with the collagen group ( P = .863). The collagen group showed less bone formation than the amnion group ( P = .194), but this difference was not significant. However, bone formation in the amnion group was significantly more than in the control group ( P = .050). Using the amniotic membrane appears to accelerate bone formation in guided bone regeneration. However, further studies should investigate its clinical impact on bone healing.

Comparative Evaluation of Coronally Advanced Flap Using Amniotic Membrane and Platelet-rich Fibrin Membrane in Gingival Recession: An 18-Month Clinical Study.

Background:An amnion membrane is a placenta-derived tissue that consists of numerous growth factors, proteins, and stem cell reserves which help in accelerated wound healing and regeneration. Platelet-rich fibrin (PRF) also releases growth factors after activation from the platelets and gets trapped within fibrin matrix which has been shown to stimulate the mitogenic response in the periosteum for bone repair and regeneration during normal wound healing. This preliminary, controlled, randomized clinical trial with an 18-month follow-up was aimed to evaluate the effectiveness of coronally advanced flap (CAF) with either PRF membrane or bioresorbable amniotic membrane (AM) in treatment of localized gingival recession defects.Materials and Methods:Sixteen healthy adult patients presenting with Miller Class I recession defects were treated surgically with CAF along with AM (Group I) or PRF (Group II) for coverage of the recession defects. For all patients, plaque index, gingival index, bleeding on probing, clinical attachment level, depth of recession, width of recession, width of attached gingiva, and gingival thickness were evaluated at 6 months and 18 months postoperatively. Statistical analysis was done using paired t-test, repeated measure analysis of variance test, Bonferroni test for intragroup comparison and unpaired t-test for intergroup comparison.Results:The results showed statistically nonsignificant (P < 0.01) difference in all clinical parameters at the 6- and 18-month follow-ups in both groups. Gingival recession in both PRF and amnion group when evaluated individually, significantly reduced from baseline to 6 months (P = 0.000) and from baseline to 18 months (P = 0.000). However, the mean value from 6 months to 18 months was statistically nonsignificant.Conclusion:The present study demonstrated that both CAF + PRF and CAF + AM are equally effective in providing clinically significant outcomes with respect to root coverage with AM showing the better percentage of root coverage as compared to PRF.

Grafting with Cryopreserved Amniotic Membrane versus Conservative Wound Care in Treatment of Pressure Ulcers: A Randomized Clinical Trial

To compare the healing process of pressure ulcers treated with cryopreserved human amniotic membrane allograft and routine pressure ulcer care in our hospital.From January 2012 to December 2013, in a prospective randomized clinical trial (IRCT201612041335N2), 24 patients with second and third stage of pressure ulcers were enrolled in this study. All patients needed split-thickness skin grafts for pressure ulcer-wound coverage. Selected patients had symmetric ulcers on both upper and lower extremities. The patients were randomly divided into two groups: amnion and control. In the amnion group, the ulcer was covered with cryopreserved amniotic membrane and in the control group it was treated with local Dilantin powder application. The duration and success rate of complete healing was compared between the two groups.The study group was composed of 24 pressure ulcers in 24 patients (19 males and 5 females) with a mean age of 44±12.70 years. The demographic characteristics, ulcer area, and underlying diseases were similar in both groups. The early sign of response, such as decrease in wound discharge, was detected 12-14 days after biological dressing. Complete pressure ulcer healing occurred only in the amnion group (p< 0.001). Partial healing was significantly higher in the amnion group (p< 0.03). Healing time in this group was faster than that the control group (20 days versus 54 days). No major complication was recorded with amniotic membrane dressing.Cryopreserved amniotic membrane is an effective biologic dressing that promotes re-epithelialization in pressure ulcers.

Grafting with Cryopreserved Amniotic Membrane versus Conservative Wound Care in Treatment of Pressure Ulcers: A Randomized Clinical Trial.

To compare the healing process of pressure ulcers treated with cryopreserved human amniotic membrane allograft and routine pressure ulcer care in our hospital. From January 2012 to December 2013, in a prospective randomized clinical trial (IRCT201612041335N2), 24 patients with second and third stage of pressure ulcers were enrolled in this study. All patients needed split-thickness skin grafts for pressure ulcer-wound coverage. Selected patients had symmetric ulcers on both upper and lower extremities. The patients were randomly divided into two groups: amnion and control. In the amnion group, the ulcer was covered with cryopreserved amniotic membrane and in the control group it was treated with local Dilantin powder application. The duration and success rate of complete healing was compared between the two groups. The study group was composed of 24 pressure ulcers in 24 patients (19 males and 5 females) with a mean age of 44±12.70 years. The demographic characteristics, ulcer area, and underlying diseases were similar in both groups. The early sign of response, such as decrease in wound discharge, was detected 12-14 days after biological dressing. Complete pressure ulcer healing occurred only in the amnion group (p< 0.001). Partial healing was significantly higher in the amnion group (p< 0.03). Healing time in this group was faster than that the control group (20 days versus 54 days). No major complication was recorded with amniotic membrane dressing. Cryopreserved amniotic membrane is an effective biologic dressing that promotes re-epithelialization in pressure ulcers.

Efficacy of freeze-dried amnion graft following hysteroscopic adhesiolysis of severe intrauterine adhesions.

To evaluate the efficacy of freeze-dried amnion graft for prevention of intrauterine adhesion (IUA) reformation after hysteroscopic adhesiolysis. A prospective randomized controlled trial was conducted among 88 women with severe IUAs who underwent hysteroscopic adhesiolysis at Beijing Obstetrics and Gynecology Hospital between July 15, 2015, and July 1, 2016. All participants had a balloon inserted into the uterine cavity for 1week. Sterilized freeze-dried amnion graft covered the balloon portion of the Foley catheter among patients allocated to the amnion group (n=44), whereas patients in the control group (n=44) did not receive the graft. Follow-up hysteroscopy was performed 3months after surgery. Preoperative and postoperative IUA scores, menstruation scores, and pregnancy rates were assessed. Both groups exhibited reductions in IUA scores and improvements in menstruation scores following treatment (P<0.001 for each measure). Compared with the control group, the amnion group had a lower IUA score (P=0.032) and a higher menstruation score (P<0.001) at follow-up. By contrast, the rates of IUA reformation and pregnancy were not significantly different between the two groups. Use of freeze-dried amnion graft was effective in reducing IUA reformation and improving menstruation (according to pictorial blood-loss assessment chart) following hysteroscopic adhesiolysis of severe IUAs. ClinicalTrials.gov: (NCT02496052).

Using Amniotic Membrane as a Novel Method to Reduce Post-burn Hypertrophic Scar Formation: A Prospective Follow-up Study.

Background:Several studies have shown that the application of amniotic membrane as a biological dressing in the management of burns is accompanied by rapid re-epithelialisation. In this follow-up study, we aimed to evaluate the possible role of amniotic membrane as an adjunct to split thickness skin grafting on reducing itching and severity of hypertrophic scar formation.Materials and Methods:From October 2013 to January 2015, in a prospective follow-up study, 54 patients (108 limbs) with second and third degree burns, covering 4%–15% of total body surface area (TBSA), were included in the study. All patients needed split-thickness skin grafts for burn-wound coverage. Selected patients had symmetric burns on two (upper or lower) extremities. Then, in every patient, the extremities were randomly divided into two groups: In one limb, the skin graft was traditionally fixed with skin staples (control group) and in the other limb, the skin graft was covered with an amniotic membrane (amnion group). Therefore, in every patient, the graft was covered with an amniotic membrane in one extremity and fixed with skin staples in the other extremity. Finally, after 6 months, the degree of itching and hypertrophic scar formation was compared between the two groups.Results:The study group was composed of 108 limbs in 54 patients (27 males and 27 females) with a mean age of 23.54 ± 4.9 years and burn 9.03 ± 2.69% TBSA. The patients were divided into two groups: 54 limbs in amnion group and 54 limbs in control group. In 59.25% of the cases, patient had less itching in the extremity covered with amniotic membrane. Furthermore, in 64.81% of the cases, patients had less hypertrophic scar formation in the extremity covered with amniotic membrane. These differences were statistically significant (P < 0.001).Conclusions:Amniotic membrane used as an adjunct in split thickness skin grafting is a novel modality which significantly reduces scar formation and itching that can be greatly distressing to burn patients. However, still more prospective well designed studies are needed to prove it.

Comparative Evaluation of Platelet Rich Fibrin and Dehydrated Amniotic Membrane for the Treatment of Gingival Recession- A Clinical Study.

The main objective of any therapeutic intervention aimed at root coverage is to restore the tissue margin to the cemento-enamel junction and to regenerate the lost periodontium. To compare the effectiveness of Platelet Rich Fibrin (PRF) and Amniotic Membrane (AM) in the treatment of gingival recession by Coronally Advanced Flap (CAF) Technique. Thirty systemically healthy patients who met inclusion-exclusion criteria were selected. After Phase-I therapy, patients were randomly allocated to one of the groups; PRF (Group-A) and Amnion Group (Group-B). Clinical parameters like Plaque Index (PI), Recession Depth (RD) and Width of Keratinized Gingiva (WKG) were measured on the baseline day, three months and six months postoperatively. Mann-Whitney test and Wilcoxon signed ranks test were applied for intergroup and intragroup comparisons respectively. Intergroup comparison between platelet rich fibrin and amniotic membrane showed statistically insignificant difference in the recession depth and width of keratinized gingiva at three and six months postoperatively. Both the membranes were equally effective in terms of recession coverage and increase in width of keratinized gingiva.

Efficacy of human amniotic membrane and collagen in maxillofacial soft tissue defects – A comparative clinical study

ObjectiveManagement of traumatic facial soft tissue injury poses a major challenge for surgeons due to aesthetic concerns. Human amniotic membrane (AM) and collagen membrane (CM), biologic materials, have attained importance in various clinical fields, especially in wound healing. Though both materials have their own unique properties, there is a need to compare and evaluate the efficacy of AM and CM as a wound dressing material for post-traumatic soft tissue injuries of the maxillofacial region. MethodsA study was conducted in Department of Oral And Maxillofacial Surgery, at our institution which encompassed all the patients (total of 20 patients) with facial soft tissue injuries (abrasions), due to road traffic accidents from January 2014 to March 2014. They were randomly divided into two groups, with 10 patients treated with AM and 10 patients treated with CM. The groups were compared according to the wound compliance, time taken for granulation tissue formation and scarring. ResultsIt was observed that the pain score was much higher in Collagen group than in Amnion group and complete adherence of the membrane to the wound was detected in both the groups. A statistically significant result in favour of AM was found with regard to average time taken for granulation tissue formation (P=0.036) and the resultant scarring. Incidence of wound infection was reported in two cases in Collagen group. ConclusionWhen compared to Collagen, AM had good adhesive, antibacterial, anti-immunogenic and anti-scarring property. Its low cost of procurement and storage makes it an ideal wound dressing material.

Trabeculectomy combined with implantation of amniotic membrane or glaucoma drainpipe implants for glaucoma

Objective To study the clinical effects and complications of the trabeculectomy combined with implantation of biologic amniotic membrane or glaucoma drainpipe implants for the treatment of primary glaucoma. Methods One hundred and fifty eyes of 125 cases with primary glaucoma were randomly divided into three groups. Patients in the amnion group (54 eyes of 46 cases) were treated with ordinary trabeculectomy combined with biologic amniotic membrane, Patients in the drainage group (52 eyes of 40 cases) were treated with trabeculectomy combined with glaucoma drainage implants, and patients in control group(44 eyes of 39 cases) were treated with ordinary trabeculectomy. Patients were followed up for one year. The post-operative intraocular pressure (IOP), filtration bleb and complications were observed. Results One week after operation, IOP of the amnion group was (13.76±0.81) mmHg(1 mmHg=0.133 kPa), IOP of the drainage group was (12.54±0.47)mmHg, IOP of the control group was (14.45±0.37)mmHg and the difference was not statistically significant(F=1.07, P=0.22). One year after operation, IOP of the amnion group was (11.81±0.74)mmHg, IOP of the drainage group was (11.75±0.25)mmHg and the difference was not statistically significant (P=0.06). IOP of the control group was (14.90±0.62)mmHg which was higher than that of the other two groups and the difference was statistically signicant compared with that of the other two groups (F=0.01, P=0.04). One week after operation, shallow anterior chamber happened in 6 eyes in the amnion group, 7 eyes in the drainage group and 5 eyes in control group. One year after operation, the percentage of functional filtration bleb was 92.6% in the amnion group, 94.2% in the drainage group and 65.9% in control group respectively. Conclusion The trabeculectomy combined with biologic amniotic membrane or glaucoma drainpipe implants can reduce the IOP safely and effectively, and inhibit filtering bleb cicatrization. Key words: Glaucoma, primary; Trabeculectomy; Amnion, biologic; Drainpipe, glaucoma

Enhancement of tooth eruption by using amniotic stem cells (Immunohistochemical study of VEGF marker)

Background: Tooth eruption is a localized process in the jaws which exhibits precise timing and bilateral symmetry. Develop within the jaws and their eruption is a complex infancy process during which they move through bone to their functional positions within the oral cavity. For species with more than one set of teeth, eruption of the second set also accomplishes. The key to the successful clinical management of tooth eruption consists of understanding that this process consists largely of the local regulation of alveolar bone metabolism to produce bone resorption in the direction of eruption and shift and formation of bone at the opposite side.The amniotic sac contains a considerable quantity of stem cells. These amniotic stem cells are able to differentiate into various tissues, which used in many field. Vascular endothelial growth factor (VEGF) is an important angiogenic factor reported to induce migration and proliferation of endothelial cells, enhance vascular permeability, and modulate thrombogenicity. VEGF expression in cultured cells (smooth muscle cells, macrophages, endothelial cells) is controlled by growth factors and cytokines. The aim of this study was to study the administration of cell molecules of (Chorion, Amnion and Amniotic fluid) around developing mouse tooth and studying the expression of VEGF marker. Materials and Methods: forty eight albino Swiss mice of one day old age injected with isolated amniotic stem cells in the anterior region of maxilla (incisors area) other 16 mice injected with saline represents control. Sacrifice 4 mice for each period (4, 7, 10, and 13) day old age. The result were studied histologically and immunohistochemistry. Results: VEGF marker localized and identified in 3 areas; pulp, P.D.L, and Bone. In pulp. The mean value of positive VEGF expression showed to be highest in Amnion group in comparison to the other studied groups. The marginal mean value of all periods reported to be highest in Amnion groups followed by Chorion group. The period 10 day showed highest marginal means value for positive VEGF expression for all groups. In P.D.L. area Amniotic fluid records the highest mean and marginal mean value specifically at day-10 in comparison to other studied groups. In Bone area Amniotic fluid records the highest mean and marginal mean value among the studied groups followed by Chorion group. Period 7-day and 10-day shows high mean value for VEGF expression. Coincidence test for VEGF marker illustrates to be affected by Amniotic fluid application in P.D.L. and in bone area while Amnion and Chorion application showed to be concerned with pulp. Conclusion. It reported that amniotic fluid application affected on expression of VEGF in P.D.L and bone while amnion and chorion showed to affect on expression of VEGF in pulp.The present study highlighted on clinical and researcher application of Amniotic fluid and Chorion for supplement of stem cell in dental tissue engineering or even in other body tissues.

Efficacy assessment of drug-loaded amniotic membrane transplantation for infectious corneal ulceration

Background Treatment of corneal ulceration by transplanting drug-loaded amniotic membrane has been used widespreadly abroad,however,seldom study is found in China up to now.Objective This study was to explore the sustained release property and the efficacy of the drug-loaded amniotic membrane.Methods The bacteriostatic area of amniotic membrane fragments immersed with different concentrations (5,20,30 g/L) of levofloxacin for different time points (5,15,30,60 minutes) was evaluated by in vitro test.Bacterial corneal ulceration models were established in 20 rabbits by injected 0.7 MCF staphylococcus aureus suspension (0.1 ml) into the central corneal stroma to form the cloudy area of 4.0-6.0 mm.Then the rabbits were randomized into two groups.Regular amniotic membrane transplantation was performed laterally and 0.5％ levofloxacin drops was topically administered after operation in the amnion+levofloxacin drops group,and drug-loading amnion transplantation was used in the drug-loading amnion group.Aqueous humor of 0.1 ml was collected in 30 minutes,1 hour,2,3.5,5.5 hours after levofloxacin was administered and 1 day,3,7,10,14,21 days after operation for the detect of levofloxacin level with high-performance liquid chromatography.The corneal symptom was scored based on McNeill's criteria in 1 week,2 weeks and 4 weeks and the ulceration area was assessed under the slit lamp in the first week.The pathological examination was carried out in the fourth week after surgery.Results The mean bacteriostatic area was bigger with the increase of levofloxacin concentration,and bacteriostatic area in amnion immersed for 15 minutes was bigger than that of 5 minutes (P＜0.01).The levofloxacin concentration of aqueous humor after transplantation was decreased by extending the time,and that in 30 minutes and 5.5 hours after operation was (0.873±0.264) mg/L and (0.106±0.027) mg/L,respectively,in the amnion+levofloxacin drops group,and that in day 1,3,7 after surgery in the drug-loading amnion group was higher than at 30 minutes in the amnion+levofloxacin drops group,showing all significant differences (all P =0.00).In the first,second and fourth week after operation,the corneal symptom score was 1.7±0.6,1.3±0.5,0.2±0.4 in the drug-loading amnion group and 2.2±0.8,2.0±0.6,1.5±0.8 in the amnion+ levofloxacin drops group,with the significant differences among the different groups and time points (Fgroup =9.49,P =0.01 ;Ftime =22.96,P=0.01).The corneal ulceration area was (1.6±0.6) mm2 in the drug-loading amnion group and (3.2±0.8) mm2 in the amnion+levofloxacin drops group 1 week after operation,showing a significant difference between them (t =3.98,P =0.00).Histopathological revealed that the various layers of cornea tissue appeared irregular arrangement in the amnion + levofloxacin drops group 4 weeks after operation with 1-2 layers of new squamous epithelium.Disorder hypothallus structure,more inflammatory cells and residual vascular cavity were visible.However,new squamous epithelium of 4-5 layers was seen in the drug-loading amnion group,and inflammatory cells and residual vascular cavity were less than the amnion+levofloxacin drops group 4 weeks after operation.Conclusions Levofloxacin-loaded amniotic membrane can sustained release levofloxacin and maintain an effective drug concentration in aqueous humor,which improves the treating efficacy for staphylococcus aureus corneal ulceration. Key words: Sustained release; Amniotic membrane/drug-loading amnion; Corneal ulceration; Levofloxacin; Ocular surface reconstruction

Effect of amniotic membrane on graft take in extremity burns

BackgroundSeveral studies have shown that the application of amniotic membrane as a biological dressing in the management of burns is accompanied by rapid re-epithelialisation and healing as it diminishes the oozing of plasma, bacterial count and fluid, protein and heat loss. This study evaluates the effect of amniotic membrane on graft take in split-thickness skin graft of extremity burns. MethodsFrom October 2008 to January 2010, in a prospective clinical trial, 54 patients (108 limbs) with second and third degree burns, covering 4–15% of total body surface area (TBSA), were included in this study. All patients needed split-thickness skin grafts for burn-wound coverage. Selected patients had symmetric burns on two (upper or lower) extremities. Then in every patient, the extremities were randomly divided into two groups: in one limb, the skin graft was traditionally fixed with skin staples (control group) and in the other limb the skin graft was covered with an amniotic membrane (amnion group). Therefore, in every patient the graft was covered with an amniotic membrane in one extremity and fixed with skin staples in the other extremity. Finally, the duration and success rate of complete graft take was compared between the two groups. ResultsThe study group was composed of 108 limbs in 54 patients (27 males and 27 females) with a mean age of 23.54±4.9 years and burn 9.03±2.69% TBSA. The mechanism of burn was flame (63%), scald (18.5%) and flash (18.5%). The rate of complete graft take was 96.76% and 88.79% in the amnion group and in the control group, respectively. The mean duration of graft take was 6.98±1.35 days in the amnion group and 13.9±1.66 days in the control group. This difference was statistically significant (P<0.001). ConclusionsOur results show that although the amniotic membrane has no negative impact on graft take, it significantly reduces the duration of complete graft take, which is very important for both the patient and the health-care system.

Efficacy of amniotic membrane-assisted bleb revision for elevated intraocular pressure after filtering surgery

To compare the effectiveness of standard revision surgery using mitomycin C (MMC) with revision using amniotic membrane transplantation and MMC for elevated intraocular pressure (IOP) after trabeculectomy. A retrospective, nonrandomized comparative study of 36 eyes of 36 patients with a failed trabeculectomy. Patients were divided into two groups, ie, a nonamnion-transplanted group and an amnion-transplanted group. The amniotic membrane was placed on the scleral flap under the conjunctiva in the amnion-transplanted group. Both groups recovered filtration of aqueous humor from the surgical site with the adjunctive use of MMC. The changes in IOP and cumulative survival rate were compared for the two groups. Success was defined as a 30% reduction in IOP from the preoperative IOP and maintenance below 21 mmHg with or without the use of antiglaucomatous agents. The mean preoperative IOP was not significantly different in the two groups. The mean postoperative IOP in the nonamnion group, 12.1 +/- 5.5 mmHg, was significantly lower than the IOP in the amnion group, 16.0 +/- 3.7 mmHg. Survival curves in the two groups did not reach significantly different levels. Conventional surgical bleb revision with MMC can significantly reduce the elevated IOP associated with a failed filtration bleb. The use of an amniotic membrane transplant did not improve the surgical outcome in our cases.