器官衰竭仍然是導致全球死亡的一個問題。然而，器官捐贈的急
劇減少加劇了全球可移植器官的短缺。因此，脫細胞支架的概念已被
選為組織工程和器官置換的臨床策略新概念。將天然組織的去細胞化
製成生物支架或生醫材料是一種很有前途的技術，可以在組織工程和
再生醫學中提供大量醫材來源。另一方面，它仍然存在各種限制，例
如未清除的殘留物會誘導免疫反應，並可能在移植後導致嚴重的炎症
反應。本研究旨為利用豬膀胱製備異種脫細胞之細胞外基質 (dECM) 支
架，以探索血清對脫細胞過程的影響。
本實驗將豬膀胱分為四組進行脫細胞程序，在原本使用去污劑、
十二烷基硫酸鈉 (SDS) 的過程中，分別添加胎牛血清、豬血清和馬血清
等三種不同類型的血清，並在一個完整的脫細胞過程後，確認脫細胞
功效之評估。藉由在dECM 的水凝膠中進行細胞培養，以評估dECM
的水凝膠之生物活性和炎症反應。由實驗結果可得知，只有用SDS 單
一處理的組別，有大量DNA 殘留，並與未脫細胞組織沒有顯著差異；
然而，在經血清處理後能降低DNA 殘留和 α-gal 表位含量減少。其中
在添加胎牛血清組別所製備的dECM 之 DNA 殘留量為 15.71 ng/mg，而
豬血清和馬血清製備的dECM之DNA殘留量分別為 44.9 和 46.82 ng/mg，
這三個組別的dECM中DNA殘留量皆低於產生免疫反應的50 ng/mg建
議值。此外與天然組織相比，這三組dECM 在免疫組織化學中顯示了
α-gal 被有效去除。此外，當與 RAW 264.7 和 THP-1 細胞與未處理的
水膠一起培養時，會刺激水凝膠釋放 TNF-α（一種促炎細胞因子），
II
然而在脫細胞中加入血清組別的水膠卻顯示能降低 TNF-α 的分泌，顯
示其較不會引起發炎反應。綜合上述，本研究結果顯示：在脫細胞步
驟中，加入10-20%之血清可以有效減少豬膀胱dECM 上的DNA 和 α-
gal。利用此脫細胞技術所製備的豬膀dECM 是一種有高度生物相容性
的醫材，並且有潛力成為應用在細胞培養和移植之免疫調節材料。

Organ failure remains a concern leading to mortality across the globe. However, the
steep reduction in organ donation aggravates the worldwide shortage of transplantable
organs. Hence, the concept of the decellularized scaffold has been chosen as a novel for
tissue engineering and clinical strategy for organ replacement. On one hand, the
decellularization of native tissue is well known as a promising technique that can offer
numerous applications in tissue engineering and regenerative medicine. On the other hand,
there are still various limitations like remnants of materials that can induce an immune
response and may lead to a severe inflammatory response upon transplantation. This study
aims to fabricate a xenogeneic decellularized extracellular matrix (ECM) scaffold from the
porcine bladder to explore the effect of serums on the decellularization process.
The decellularization procedure was developed and porcine bladders were divided
into four groups and deflated by detergent, Sodium Dodecyl Sulfate (SDS) and its
modification, using three different types of serum, Fetal calf serum, Porcine serum, and
Horse serum under temperature control. After a complete process, the evaluation of
decellularization efficacy was confirmed. Consequently, dECM-based hydrogel
bioscaffold and cell seeding were fabricated in sterilized conditions to assess the bioactivity
and inflammatory response. All samples resulted in a reduction of recombinant DNA and
alpha-gal epitope content on each sera treatment. Only the group of single treatment with
SDS showed a high content of DNA with no significantly different from undecellularized
tissue. Nevertheless, the result on each sera treatment indicated a significant decrease of
DNA, with 15.71 ng per mg dry weight ECM left after incubating in fetal calf serum, 44.9,
IV
and 46.82 ng/mg on porcine serum, and horse serum, respectively. Immunohistochemistry
also showed the removal of alpha-gal epitopes compared to the native tissues. In addition,
quantification of released TNF-α (a pro-inflammatory cytokine) stimulated by hydrogel
when cultured with RAW 264.7 and THP-1 cell line revealed no immunological response
(TNF-alpha secretion inhibits). Inclusively, our findings imply that serum can reduce both
nuclease and alpha-gal epitopes on the porcine bladder so that the acellular porcine bladder
can be considered a biocompatible material, and is suitable for cell culture and
transplantation applications for immunomodulatory materials.

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