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异体脂肪和筋膜基质的解构:筋膜基质在啮齿动物模型中提高血管生成、体积保持和脂肪生成。

Deconstructing Allograft Adipose and Fascia Matrix: Fascia Matrix Improves Angiogenesis, Volume Retention, and Adipogenesis in a Rodent Model.

机构信息

From the Center for Tissue Engineering, Department of Plastic Surgery.

MTF Biologics.

出版信息

Plast Reconstr Surg. 2023 Jan 1;151(1):108-117. doi: 10.1097/PRS.0000000000009794. Epub 2022 Oct 11.

Abstract

BACKGROUND

Autologous fat grafting is commonly used for soft-tissue repair (approximately 90,000 cases per year in the United States), but outcomes are limited by volume loss (20% to 80%) over time. Human allograft adipose matrix (AAM) stimulates de novo adipogenesis in vivo, but retention requires optimization. The extracellular matrix derived from superficial fascia, interstitial within the adipose layer, is typically removed during AAM processing. Thus, fascia, which contains numerous important proteins, might cooperate with AAM to stimulate de novo adipogenesis, improving long-term retention compared to AAM alone.

METHODS

Human AAM and fascia matrix proteins (back and upper leg regions) were identified by mass spectrometry and annotated by gene ontology. A three-dimensional in vitro angiogenesis assay was performed. Finally, AAM and/or fascia (1 mL) was implanted into 6- to 8-week-old male Fischer rats. After 8 weeks, the authors assessed graft retention by gas pycnometry and angiogenesis (CD31) and adipocyte counts (hematoxylin and eosin) histologically.

RESULTS

Gene ontology annotation revealed an angiogenic enrichment pattern unique to the fascia, including lactadherin, collagen alpha-3(V) chain, and tenascin-C. In vitro, AAM stimulated 1.0 ± 0.17 angiogenic sprouts per bead. The addition of fascia matrix increased sprouting by 88% (2.0 ± 0.12; P < 0.001). A similar angiogenic response (CD31) was observed in vivo. Graft retention volume was 25% (0.25 ± 0.13) for AAM, significantly increasing to 60% (0.60 ± 0.14) for AAM/fascia ( P < 0.05). De novo adipogenesis was 12% (12.4 ± 7.4) for AAM, significantly increasing to 51% (51.2 ± 8.0) for AAM/fascia ( P < 0.001) by means of adipocyte quantification.

CONCLUSIONS

Combining fascia matrix with AAM improves angiogenesis and adipogenesis compared to AAM alone in rats. These preliminary in vitro and pilot animal studies should be further validated before definitive clinical adoption.

CLINICAL RELEVANCE STATEMENT

When producing an off-the-shelf adipose inducing product by adding a connective tissue fascial component (that is normally discarded) to the mix of adipose matrix, vasculogenesis is increased and, thus, adipogenesis and graft survival is improved. This is a significant advance in this line of product.

摘要

背景

自体脂肪移植常用于软组织修复(美国每年约有 9 万例),但随着时间的推移,其体积损失(20%至 80%)会导致效果受限。同种异体脂肪基质(AAM)可刺激体内新生脂肪形成,但保留率需要优化。来源于浅筋膜的细胞外基质,即脂肪层内的间质,在 AAM 处理过程中通常被去除。因此,筋膜可能含有许多重要的蛋白质,它可能与 AAM 合作刺激新的脂肪形成,与单独使用 AAM 相比,可改善长期保留率。

方法

通过质谱法鉴定人 AAM 和筋膜基质蛋白(背部和大腿区域),并通过基因本体论进行注释。进行了三维体外血管生成分析。最后,将 AAM 和/或筋膜(1ml)植入 6-8 周龄雄性 Fischer 大鼠体内。8 周后,作者通过气体比重计评估移植物保留情况,并通过 CD31 和脂肪细胞计数(苏木精和伊红)进行组织学评估血管生成和脂肪形成。

结果

基因本体论注释显示,筋膜具有独特的血管生成富集模式,包括乳白蛋白、α-3(V) 链胶原和 tenascin-C。体外,AAM 刺激每个珠子产生 1.0±0.17 个血管生成芽。添加筋膜基质可使芽的生成增加 88%(2.0±0.12;P<0.001)。体内也观察到了类似的血管生成反应(CD31)。AAM 的移植物保留量为 25%(0.25±0.13),而 AAM/筋膜的移植物保留量显著增加至 60%(0.60±0.14)(P<0.05)。AAM 的新生脂肪形成率为 12%(12.4±7.4),而 AAM/筋膜的新生脂肪形成率显著增加至 51%(51.2±8.0)(P<0.001),通过脂肪细胞计数进行评估。

结论

与单独使用 AAM 相比,将筋膜基质与 AAM 结合可提高血管生成和脂肪形成,这在大鼠体内的初步体外和初步动物研究中得到了验证,在最终临床应用之前,还需要进一步验证。

临床意义

在通过添加结缔组织筋膜成分(通常被丢弃)来混合脂肪基质来生产现成的脂肪诱导产品时,血管生成增加,因此脂肪形成和移植物存活率提高。这是该产品的一个重大进展。

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