Laboratório de Ginecologia Estrutural e Molecular (LIM-58), Departamento de Obstetrícia e Ginecologia, Hospital das Clínicas da Faculdade de Medicina da Universidade de São Paulo, São Paulo, SP, Brazil.
Laboratório de Ginecologia Estrutural e Molecular (LIM-58), Departamento de Obstetrícia e Ginecologia, Hospital das Clínicas da Faculdade de Medicina da Universidade de São Paulo, São Paulo, SP, Brazil.
Clinics (Sao Paulo). 2022 Jun 28;77:100066. doi: 10.1016/j.clinsp.2022.100066. eCollection 2022.
Gelfoam scaffold is a feasible and safe non-invasive technique for Adipose tissue-derived Stem Cell (ASC)-delivery in the treatment of frozen-thawed ovarian autografts. This study seeks to analyze the genes expression profile of rat frozen-thawed ovarian autografts treated with scaffold-based delivery of adipose tissue-derived stem cells.
Eighteen adult Wistar rats were distributed into three groups: Control (frozen-thawed only); Group 1 (G1) and Group 2 (G2) (frozen-thawed ovaries treated with culture medium or ASC, respectively). Both treatments were performed immediately after autologous retroperitoneal transplant with scaffold-based delivery. The ovarian grafts were retrieved 30 days after transplantation. Quantitative gene expression (qPCR) for apoptosis, angiogenesis, and inflammatory cytokines (84 genes in each pathway) were evaluated by RT-PCR. Graft morphology (HE), apoptosis (cleaved-caspase-3), neoangiogenesis (VEGF), and cellular proliferation (Ki-67) were assessed.
In grafts treated with ASC, the apoptosis pathway showed the highest number of genes over-regulated - 49 genes - compared to inflammation cytokines and angiogenesis pathway - 36 and 23 genes respectively, compared to grafts treated with culture medium. Serpinb5 family was highlighted in the angiogenesis pathway and Cxcl6 in the inflammation cytokines pathway. In the apoptosis pathway, the most over-regulated gene was Capsase14. ASC treatment promoted the reduction of cleaved caspase-3 in the theca internal layer and increased cell proliferation by Ki-67 in the granulosa layer without altering VEGF. A mild inflammatory infiltrate was observed in both groups.
ASC therapy in rat frozen-thawed ovarian autografts promoted an abundance of genes involved with apoptosis and inflammatory cytokines without compromising the ovary graft morphology and viability for short time. Further studies are necessary to evaluate the repercussion of apoptosis and inflammation on the graft in the long term.
Gelfoam 支架是一种可行且安全的非侵入性技术，可用于将脂肪组织源性干细胞（ASC）递送至冷冻解冻的卵巢自体移植物中。本研究旨在分析接受支架递送脂肪组织源性干细胞治疗的冷冻解冻卵巢自体移植物的基因表达谱。
将 18 只成年 Wistar 大鼠随机分为三组：对照组（仅冷冻解冻）；第 1 组（G1）和第 2 组（G2）（分别用培养基或 ASC 处理冷冻解冻的卵巢）。两种处理均在支架辅助逆行腹膜后移植后立即进行。移植后 30 天取回卵巢移植物。通过 RT-PCR 评估细胞凋亡、血管生成和炎症细胞因子（每条途径 84 个基因）的定量基因表达（qPCR）。评估移植物形态（HE）、凋亡（cleaved-caspase-3）、新生血管（VEGF）和细胞增殖（Ki-67）。
与用培养基处理的移植物相比，用 ASC 处理的移植物中，凋亡途径的基因上调数量最多 - 49 个基因 - 其次是炎症细胞因子和血管生成途径 - 分别为 36 和 23 个基因。Serpinb5 家族在血管生成途径中突出，Cxcl6 在炎症细胞因子途径中突出。在凋亡途径中，上调最明显的基因是 Capsase14。ASC 治疗促进了内层颗粒细胞层中 cleaved caspase-3 的减少，并通过 Ki-67 增加了颗粒细胞层的细胞增殖，而不改变 VEGF。两组均观察到轻度炎症浸润。
在大鼠冷冻解冻卵巢自体移植物中，ASC 治疗促进了大量与细胞凋亡和炎症细胞因子相关的基因的表达，而不会在短期内损害卵巢移植物的形态和活力。需要进一步研究来评估细胞凋亡和炎症对移植物的长期影响。
