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Published on: May 17, 2015
Quantitative analysis of host cells growing into canine homograft valved aortic and pulmonary artery
Jian-hua Yu1, Hong-wei Guo, Shi-qiu Song
1Department of Cardiovascular Surgery, Qilu Hospital, Shandong University, Jinan, Shandong 250012, China. yujianhua1965@126.com
Insights
Host cell infiltration is crucial for the long-term function of cryopreserved conduit valved homografts (CVH). Storage methods (4°C vs. liquid nitrogen) did not significantly impact host cell ingrowth or graft morphology.
Area of Science:
- Cardiovascular surgery
- Tissue engineering
- Cryobiology
Background:
- Cryopreserved conduit valved homografts (CVH) are utilized in cardiac surgery.
- Assessing host cell integration, durability, and immunogenicity is vital.
- Comparing 4°C storage versus liquid nitrogen cryopreservation for CVH performance.
Purpose of the Study:
- To quantify host cell ingrowth in CVH.
- To evaluate the durability and immunogenicity of CVH.
- To compare CVH stored at 4°C with those cryopreserved in liquid nitrogen.
Main Methods:
- Heterotopic canine CVH transplantation (n=14 per group).
- Quantitative PCR for donor/host cell ratios.
- Immunohistochemistry (FGF-2) and electron microscopy for tissue viability.
Main Results:
- No significant difference in host cell ingrowth or morphology between storage methods.
- Host cell infiltration increased over 12 months, reaching 40-47%.
- Fibroblasts and endothelial cells were primary invading cell types.
Conclusions:
- Host cell ingrowth is essential for CVH structural and functional integrity.
- Both 4°C and liquid nitrogen storage yield comparable outcomes for CVH allografts.
Background:
Cryopreserved conduit valved homografts (CVH) have been widely used in surgical treatment of cardiac disease. This study aimed to determine the extent of host cell ingrowth and the durability and immunogenicity of CVH, and to compare the performance of CVH stored at 4°C and CVH cryopreserved in liquid nitrogen at -196°C.
Methods:
Heterotopic transplants of canine CVH stored at 4°C (n = 14) and cryopreserved in liquid nitrogen (n = 14) were made onto the abdominal aorta of recipient dogs. Animals were sacrificed at 7 and 15 days and at 1, 3, 6, 9, and 12 months after transplantation to excise the implanted CVHs. Tissue DNA extraction and quantitative polymerase chain reaction (PCR) were performed to calculate the ratio of donor cells and host cells in the CVH. The tissue viability of CVH after implantation was analyzed by detecting alkaline fibroblast growth factor 2 (FGF-2) using immunohistochemical staining and by observation under transmission electron microscope and scanning electron microscope.
Results:
All the animals survived and recovered well. There were few repopulating host cells (0.04% - 0.83%) in the implanted CVH at 7 or 15 days. The ratio of ingrowing host cells into the CVH continued rising after implantation and reached 40% - 47% in the 12th month postoperation. Histology, transmission electron microscopy and FGF-2 immunohistochemical staining indicated that fibroblasts and the host's endothelial cells were the main cellular elements invading the CVH. There were no significant differences in results between CVH stored at 4°C and CVH cryopreserved in liquid nitrogen.
Conclusions:
Host cells growing into CVH are very important for maintaining the long-term structure and function of the implanted CVH. There is no significant difference between CVH storing at 4°C or in liquid nitrogen in regard to the ingrowth of host cells or of morphologic features after CVH allografting.
