PMID- 35959605 OWN - NLM STAT- MEDLINE DCOM- 20220815 LR - 20220816 IS - 0412-4081 (Print) IS - 0412-4081 (Linking) VI - 58 IP - 8 DP - 2022 Aug 11 TI - [Therapeutic effect of amniotic membrane-fibrin sealant cement on severe ocular surface alkali burn in rabbits]. PG - 606-614 LID - 10.3760/cma.j.cn112142-20220310-00103 [doi] AB - Objective To prepare a biologically active amniotic membrane powder and explore its preservation conditions, and to evaluate the efficacy of the amniotic membrane (AM)-fibrin sealant (FS) cement made from the amniotic powder on the rabbit severe ocular surface alkali burn model. Methods Experimental research. Fresh AM was air-dried, cooled with liquid nitrogen, ground into amniotic powder and sterilized by radiation. The expression of transformed growth factor, nerve growth factor receptor (NGFR), hepatocyte growth factor (HGF), epidermal growth factor (EGF) and basic fibroblast growth factor (bFGF) after preparation and 10, 20 and 30 days after storage at room temperature, 4 ℃ and -20 ℃ was tested and compared with that in the fresh AM. The AM-FS cement containing different concentrations of amniotic powder and no amniotic powder was diluted. Rabbit corneal epithelial cells were cultured for 72 hours. The effects of different concentrations of amniotic powder on epithelial cell growth were observed by light microscopy, and the amniotic powder concentration with the largest absorbance value at 450 nm was selected for subsequent animal experiments. Thirty-two right eyes of 32 rabbits as the severe ocular surface alkali burn model were divided using the random counting method into the AM-FS cement group, fresh AM transplantation group, FS group and antibiotic control group (8 rabbits each group) and given different interventions. After weekly observation of corneal repair, hematoxylin and eosin staining, immunohistochemical staining of monocyte chemotaxis protein 1 (MCP-1)and vascular endothelial growth factor (VEGF) were performed and detected by light microscopy at 28 days. The logFC values of the growth factor or receptor expression difference ratio were corrected by BH; the data were analyzed by t-test and analysis of variance. Results: The expression of TGF in the amniotic membrane powder compared with the fresh amniotic membrane group (logFC=-0.11), and the expression of NGFR (HGF, EGF, bFGF) was higher than that of the fresh amniotic membrane group (logFC=-2.07, 0.72, 0.46, 2.62; P<0.05); the expression of HGF, bFGF and EGF in amniotic membrane powder stored for 10 days and 20 days were no lower than fresh amniotic membrane; at 30 days, the expression of growth factors or receptors except HGF and bFGF were decreased, and HGF, bFGF and EGF were no less than 4 ℃ and -20 ℃.The maximum A value was obtained for 0.25 mg/ml of the amniotic membrane powder after 72 hours of the CEC culture 0.98+/-0.05. The corneal recovery was better in the AM-FS and fresh amniotic membrane transplant groups, with corneal turbidity scores of 3.75+/-0.46 and 3.50+/-0.46, respectively, on 28 days, lower than antibiotics (4.29+/-0.45) (t=2.480, 3.629; P=0.019, 0.001). The corneal neovascular area in the antibiotic control group was compared with the other three groups (t=4.040, 4.339, 2.820; all P<0.001); the corneal neovascular area in the AM-FS group was (9.88+/-0.20) and (18.96+/-0.18) mm(2) at 7 and 28 days. The corneal neovascularization area at 7 and 28 days in the fresh AM group [(9.54+/-0.22) and (18.08+/-0.96) mm(2)] was smaller than the AM-FS group (t=3.085, 3.017, P=0.005, 0.005). Despite the tiny statistical difference (0.34, 0.88), there was no clinical difference. Hematoxylin and eosin staining showed corneal structures were intact in the AM-FS and fresh AM groups, the epithelial arrangement became normal, and the corneal healing was superior to the FS and antibiotic control groups. Immunohistochemistry showed that the positive expression of VEGF in the fresh AM group was weaker than that in the remaining three groups. MCP-1 was expressed to a similar extent in the AM-FS and fresh AM groups. Conclusions: The active cytokine had high expression and stable properties at room temperature. The AM-FS cement containing 0.25 mg/ml amniotic powder can promote the repair of corneal epithelium, reduce inflammatory reaction and corneal neovascularization after alkali burning in rabbit eyes. FAU - Li, N Y AU - Li NY AD - The Second People Hospital of Yunnan Province, Yunnan Eye Hospital, Yunnan Institute of Ophthalmology, Yunnan Key Laboratory of Ophthalmic Disease Prevention and Treatment, Yunnan Clinical Medical Center for Ophthalmic Diseases, Yunnan Clinical Medical Research Center for Eye Diseases, Kunming 650021, China. FAU - Zhang, W J AU - Zhang WJ AD - The Second People Hospital of Yunnan Province, Yunnan Eye Hospital, Yunnan Institute of Ophthalmology, Yunnan Key Laboratory of Ophthalmic Disease Prevention and Treatment, Yunnan Clinical Medical Center for Ophthalmic Diseases, Yunnan Clinical Medical Research Center for Eye Diseases, Kunming 650021, China. FAU - Hu, Z L AU - Hu ZL AD - The Second People Hospital of Yunnan Province, Yunnan Eye Hospital, Yunnan Institute of Ophthalmology, Yunnan Key Laboratory of Ophthalmic Disease Prevention and Treatment, Yunnan Clinical Medical Center for Ophthalmic Diseases, Yunnan Clinical Medical Research Center for Eye Diseases, Kunming 650021, China. LA - chi GR - 2018ZF009/Yunnan Provincial Major Science and Technology Foundation/ GR - L‑2019029/Medical Leading Talents Training Program of Yunnan Provincial Health Commission/ GR - 202001AY070001‑255/Joint Project of Applied Basic Research of Kunming Medical University, Science and Technology Department of Yunnan Province of China/ PT - Journal Article PT - Randomized Controlled Trial, Veterinary PL - China TA - Zhonghua Yan Ke Za Zhi JT - [Zhonghua yan ke za zhi] Chinese journal of ophthalmology JID - 16210540R RN - 0 (Anti-Bacterial Agents) RN - 0 (Fibrin Tissue Adhesive) RN - 0 (Powders) RN - 0 (Vascular Endothelial Growth Factor A) RN - 62229-50-9 (Epidermal Growth Factor) RN - TDQ283MPCW (Eosine Yellowish-(YS)) RN - YKM8PY2Z55 (Hematoxylin) SB - IM MH - Amnion/metabolism/transplantation MH - Animals MH - Anti-Bacterial Agents MH - *Burns, Chemical/therapy MH - *Corneal Neovascularization/metabolism MH - Eosine Yellowish-(YS)/metabolism MH - Epidermal Growth Factor/metabolism MH - Fibrin Tissue Adhesive/metabolism MH - Hematoxylin/metabolism MH - Powders/metabolism MH - Rabbits MH - Vascular Endothelial Growth Factor A/metabolism EDAT- 2022/08/13 06:00 MHDA- 2022/08/16 06:00 CRDT- 2022/08/12 04:06 PHST- 2022/08/12 04:06 [entrez] PHST- 2022/08/13 06:00 [pubmed] PHST- 2022/08/16 06:00 [medline] AID - 10.3760/cma.j.cn112142-20220310-00103 [doi] PST - ppublish SO - Zhonghua Yan Ke Za Zhi. 2022 Aug 11;58(8):606-614. doi: 10.3760/cma.j.cn112142-20220310-00103.