Objective To observe clinical effects of burn wounds treatment with bovine amnion and to screen the best method of preparing and storing of bovine amnion. Methods From January 2004 to January 2005,We selected randomly 58 patients with superficial Ⅱ° wound, deepⅡ° wound, autografting area for removal of eschars and tangential excision, fetching skin area or residual burn wound . Using auto-control, every burn wound was divided into 3 parts and was treated with 3 dressings: bovine amnion dealt with by 0.1% chlorhexidine（group A）, bovine amnion dealt with by 0.4% glutaraldehyde（group B） and vaseline gauze dressing（group C as control）. The clinical effects were compared between different groupsand the method of preparing and storing bovine amnion was evaluated. Results The dressing texture of group A was softer than that of group B, and its flexibility was fine. The pretreatment was not necessary for dressing in group A. When the dressing was used on burn wounds in groups A and B, painwas slight, but pain was obvious in group C; healing time in groups A and B was much less than that in group C, showing statistically significant difference(P＜0.01). There was no statistically significant difference in healing time between groups A and B (P＞0.05). The infection ratio of burn wound in deepⅡ° wound and residual burn wound of groups A and B is much lower than that of group C, showing statistically significant difference (P＜0.05); in theother burn wounds there was no significant difference (P＞0.05). There was no statistically significant difference between groups A and B (P＞0.05). Conclusion Bovine amnion could make benefit on burn wounds healing, reduce infection ratio of burn wounds, could be used on different kinds of burn wounds. The clinical effect between bovine amnion dealt with by glutaraldehyde and by chlorhexidine is similar. Whereas the latter is more easy to be popularized.
Objective To compare the reparative effects between the acellular small intestinal submucosa andthe acellular amnion as dressings for traumatic skin defects. Methods Three full-thickness skin defects, which wereclose to the vertebral column of the pig, were created on both sides of the dorsum. The skin defects were randomlydivided into three groups. In each group, the following different materials were used to cover the skin defects: theacellular amnion in Group A, the acellular small intestinal submucosa (SIS) in Group B, and the physiological saline aguze in Group C (the control group). The specimens from the skin defects were harvested for a histological evaluation and for determination of the hydroxyproline content at 10 (2 pigs), 20 (2 pigs), and 30 days (3 pigs). We observed the healing process of the wound and its healing rate, counted the inflammatory cells, vasecular endothelial cells, and proliferating cells, and determined the hydroxyproline content. Results The acellular amnion in Group A and acellular SIS in Group B adhered to the wound tightly, but they did not adhere to the dressing; when the dressing was changed, the wound did not bleed. The saline gauze in Group C adhred to the wound tightly, but when the dressing was changed, the wound bled until 22 days after operation. Under the microscope, the collagen in the tissue below the epithelium was arranged more regularly and there were fewer cells concerned with inflammation in Groups A and B than in Group C at 10, 20, and 30 days after operation. At 10, 20, and 30 days after operation, the wound healing rate was greater in Groups A and B than in Group C, The number of the inflammatory cells and the proliferating cells were greater in Groupo C than in Groups A and B. There was a statistically significant difference (P lt; 0.05),At 20 and 30 days after operatin, the content of hydroxyproline was greater in Group c than in Group A and B. There was a statistically significant difference (P lt; 0.05). However, there was no statistically significant difference between Group A and Group B in the wound healing rate, the numbers of the inflammatory cells, vascular endothelial cells and prokiferating cells, and the content of hydroxyproline(P gt; 0.050). There was no statistically significant difference among the three groups in the number of the vascular endothelial cells. Conclusion Compared with Group C........
ObjectiveTo evaluate the safety and efficacy of amniotic membrane patching in the treatment of recurrent macular hole associated with retinal detachment of high myopia (MHRD). MethodsA prospective study. From March 2018 to January 2020, 11 patients (11 eyes) of recurrent macular hole associated with MHRD at the First Affiliated Hospital of Zhengzhou University were enrolled. Among them, there were 3 males (3 eyes), and 8 females (8 eyes). The average age was 63.64±5.82. The axis length (AL) was 29.10±0.59 mm, and the logarithm of the minimum angle of resolution best corrected visual acuity (logMAR BCVA) was 2.23±0.57. Patients previously received pars plana vitrectomy (PPV) combined with internal limiting membrane stripping surgery, which was more than 1 time. All eyes underwent standard pars plana three-channel 23G PPV combined with amniotic membrane covering and silicone oil filling. The silicone oil was removed 6 months after surgery. Follow-up time was up to 3 months after silicone oil removal surgery. 1, 3, and 6 months after the operation, the same equipment and methods were used to conduct relevant examinations before the operation to observe the closure of the macular hole, retinal reattachment and changes in logMAR BCVA. The logMAR BCVA before and after surgery was compared by paired t test. ResultsAt 1, 3, and 6 months after the operation, the retinas of all eyes were anatomically repositioned, the macular holes were well closed, and the amniotic membrane was attached to the retina. At 3 months after the silicone oil removal operation, there was no recurrence of macular hole in all eyes; logMAR BCVA was 1.35±0.32. No serious complications occurred during and after surgery in all eyes. ConclusionAmniotic membrane patching is a safe and effective method for recurrent macular hole associated with MHRD.
ObjectiveTo observe the survival, migration, and effect of human amniotic epithelial cells (hAECs) on hepatic fibrosis in immune rats so as to provide the experimental theory for the clinical treatment with hAECs. MethodsSixty-four 10-week-old male Sprague Dawley rats (weighing, 220-280 g) were randomly divided into 4 groups, sixteen rats in each group. Rat hepatic fibrosis model was induced in groups A, B, and C; hepatic fibrosis rats were injected with 4×106 hAECs in group A, and with normal saline in group B, and no treatment was given in group C; group D served as control group. After 2 weeks of transplantation, the expression of human Alu gene repeat sequence was detected by DNA-PCR method and human leucocyte antigen G (HLA-G) by immunohistochemical staining in heart, liver, spleen, kidney, lung, and brain in group A, and then the percentage of positive expression was compared between organs except spleen. Semi-quantitative analysis was done for liver fibrosis with HE staining according to Chevallier semi-quantitative histological liver fibrosis scoring system, and immunohistochemical staining for TGF-β1 was used to record immunohistochemical score (ISH), the concentrations of aspartate transaminase (AST), alanine aminotransferase (ALT), and albumin (ALB) were determined to analyze hepatic fibrosis. ResultsAlu gene repeat sequence and HLA-G could be detected in liver, heart, brain, lung, and kidney in group A, the percentage of positive expression in the liver was significantly higher than that in the other organs (P<0.05). The histological semi-quantitative score of group A (10.47±3.20) was significantly lower than that of groups B and C[(13.84±3.46) and (13.85±3.16)](P<0.05), but no significant difference was found between groups B and C (P>0.05). The ISH scores in groups A, B, C, and D were 3.60±1.50, 5.38±2.60, 5.50±2.40, and 1.87±1.36, respectively; groups A, B, and C were significantly higher than group D, and group A was significantly lower than groups B and C (P<0.05), but there was no significant difference between groups B and C (P>0.05). The concentrations of ALT and AST in groups A, B, and C were significantly higher than those in group D, and group A was significantly lower than groups B and C (P<0.05), but there was no significant difference between groups B and C (P>0.05). The concentration of ALB in groups A, B, and C was significantly lower than that in group D, and group A was significantly higher than groups B and C (P<0.05), but there was no significant difference between groups B and C (P>0.05). ConclusionhAECs can survive in immune rats by intrasplenic transplantation and migrate to liver, heart, brain, lung, and kidney, and the liver shows the largest migration. The transplantation of hAECs in immune rat with cirrhosis can alleviate hepatic fibrosis and improve the serum indexes of liver function.
Objective To investigate the feasibility of human amniotic membrane-living skin equivalent (AM-LSE) in repairing the skin defect. Methods A 5-year-old boy with giant nevus at neck, shoulder, and back was admitted in July 2016. Normal skin tissue of the patient was harvested and keratinocytes and dermal fibroblasts were separated and expanded in vitro. Human AM was donated from a normal delivery and de-epithelialized for constructing an LSE as a matrix. Keratinocytes were seeded on the epithelial side of the AM which was previously seeded with fibroblasts on the stromal side and then the complex was lifted for air-liquid surface cultivation for 10 days and observed under naked eyes and sampled for histological study. The nevus was excised to deep fascia and the skin defect in size of 20 cm×15 cm was covered with artificial skin of collagen sponge for 2 weeks to enhance granulation tissue formation, and then the AM-LSE grafts of stamp size were grafted on. The dressing was changed until the wound healed. Results After 10 days of air-liquid surface cultivation, the AM-LSE developed a multilayered and differentiated epidermis with the fibroblasts-populated amnion as the dermal matrix. The LSE stamps survived and expanded to cover the whole wound. The grafted area showed normal skin color and soft contexture at 6 months after operation, and histological study showed well developed epidermis with compactly aligned basal cells, stratified and well differentiated squamous, granular layers and stratum corneum and well vascularized dermal compartment without inflammatory cells infiltration. Conclusion The cultivated AM-LSE with autologous cells can repair skin defect and survive for a long term without rejection.
Human amnion (hAM), as a biomaterial, has made significant progress in the field of ophthalmology, particularly in the treatment of retinal diseases. hAM possesses biological properties such as promoting tissue repair, inhibiting inflammation and neovascularization, and reducing fibrosis, which have led to its promising clinical outcomes in treating macular holes, retinal detachment, proliferative vitreoretinopathy, optic disc depression-related macular detachment, and age-related macular degeneration. The application of hAM can improve surgical success rates and promote vision recovery, with no significant rejection reactions observed due to its low immunogenicity. Nevertheless, the use of hAM still faces challenges in optimizing preparation and storage techniques, enhancing therapeutic efficacy, and reducing the risk of infectious disease transmission. Future research should focus on addressing these issues to further promote the application of hAM in retinal disease treatment and enhance its effectiveness.
Objective To observe the effect of amniotic homogenate on closing holes in experimental rhegmatogenous retinal detachment and investigate its mechanism. Methods Forty rabbits were randomly divided into group A, B, C and D with 10 rabbits in each group. Group A and C were the treatment groups, and group B and D were the control groups. All eyes of rabbits underwent pars plana vitrectomy, retinectomy, and fluidair exchange. The surface of the breaks was treated with 01 ml amniotic homogenate in experimental groups and 0.1 ml PBS in control groups. At the end of operation, 20% SF6 was tamponaded and the retina reattaced. The animals were executed 14 (group A and B) and 28 days (group C and D) after the surgery. The tissue sections were observed by light microscope, electron microscope and immunocytochemistry method. Results Fourteen days after the surgery, the retina reattached in 6 eyes in group A (60%) and 2 eyes in group B (20%) （P=0.021）. Twenty-eight days after the surgery, the retina reattached in 8 eyes in group C (80%) and 3 eyes in group D (30%) (P=0.046). The difference of the rate of retinal reattachment among the 4 groups were statistical significant (Plt;0.05). Light postoperative inflammation of ocular anterior segment was observed, which was controlled 3-5 days after treated with topical steroids. The result of light microscopy showed that the eyes in treatment groups had multilayer of fibroblastlike cells around the retinal breaks, adhering to the choroid and retinal pigment epithelial cells. The proliferative cells around the retinal breaks obvious less in control groups than that in the treatment groups, and the retina could not adhere to the choroid. The results of electron microscopy were the same as that of light microscopy. Immunohistochemistry staining of the fibroblastlike cells revealed positve glial fibrillary acidic protein, which suggested that the proliferative cells around the retinal breaks were retinal glial cells. Conclusions Amniotic homogenate helps to seal retinal breaks and promote retinal reattachment by stimulating the proliferation of retinal glial cells around the breaks.
Objective The human amniotic epithel ial cells (hAECs) are a recently identified new type of stem cells.It has previously been shown that hAECs express hepatocyte-related gene and possess intracellular features and functional properties of hepatocytes. The hAECs may be a candidate seed cell for l iver regeneration. To research the survival and migrationin vivo of hAECs via adeno-associated virus-mediated the green fluorescent protein gene (AAV-GFP) transfection, and toexplore the expression of hepatocyte-l ike function. Methods Thirty nude mice (aging 6-8 weeks, half males and females, and weighing 20-22 g) were randomly divided into 3 groups (groups A, B, and C, n=10). The mice of groups A and C were made the 2/3 partial hepatectomy model, and the mice of group B underwent open abdominal operation without hepatectomy. The hAECs transfected by AAV-GFP were transplanted into the inferior end of the spleen in groups A and B with a cell density of 5 × 106/mL and a volume of 0.2 mL; the same volume of normal sal ine was injected in group C. At 4 hours, the nude mice were sacrificed and the samples of l iver, spleen, heart, lung, brain, and kidney were harvested and the general observation, histological observation, and immunofluorescence detection were performed for the hAECs survival, migration, and the functional properties of hepatocytes. Results No tumor tissue was found in l iver and spleen of 3 groups, and HE staining showed no tumor cells. There were a lot of roundl ike and deeply-stained cells with less cytoplasm and large nucleus in the spleen and the l iver of group A; no abnormal cells were found in l iver and spleen of groups B and C and in kidney, heart, bung, and brain of groups A, B, and C. The GFP+ cells were detected in the spleen and l iver of group A with expressing human albumin, but no GFP+ cells was found in l iver and spleen of groups B and C and in heart, kidney, lung, and brain of groups A, B, and C. Conclusion AAV-GFP infected hAECs transplanted into SCID nude mice with hepatectomy can keep the hepatocyte-l ike function. It will be beneficial to further identify their biological characteristics.
Objective To study the differentiation of the human osteoblasts during the construction of the tissue engineered periosteum with the human acellular amniotic membrane(HAAM).Methods To construct the tissue engineered periosteum (n=60) with HAAM, the human fetal osteoblasts were used. The fetal osteoblasts were cultured for 2, 4, 6, 8, and10 days, and then their total RNA was extracted, which were reversely transcripted to cDNA. The realtime PCR analysis was used to reveal Cbfal and Osterix, and the cycle threshold (Ct) was also measured. The simplycultured osteoblasts were used as the control group (n=20).Results The expression of Cbfa1 was higher in the experimental group on the 2nd day when compared with that on the 4th, 6th, and 8th day(P＜0.05). The same result existed on the 10th day when compared with that on the 4th and 8th day. The expression of Osterix increased and was highest on the 8th day when compared with the other results(P＜0.05). Both of the 2 gene expressions were decreased in the control group when compared with those in the experimental group, but with no significant difference(P＞0.05). Conclusion Cbfa1 and Osterix can be normally expressed by the osteoblasts after their integration with HAAM. As a scaffold, HAAM can be used to keep the osteoblast phenotype and differentiation with an osteoconductive ability. Such a cell-scaffold complex may provide a basis for the osteogenesis.