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Counting CD4(+) and CD8(+) T cells in the spleen: a novel in vivo method for assessing biomaterial immunotoxicity
Shyh-Jou Shieh1, Prashanth Varkey1, Po-Yang Chen1
1Division of Plastic and Reconstructive Surgery, Department of Surgery, National Cheng Kung University Medical College and Hospital, Tainan, Taiwan, Research Center of Excellence in Regenerative Medicine, National Cheng Kung University, Tainan, Taiwan, Advanced Optoelectronic Technology Center, National Cheng Kung University, Tainan, Taiwan, International Research Center for Wound Repair and Regeneration (iWRR), National Cheng Kung University, Tainan, Taiwan, Institute of Biotechnology, College of Bioscience and Biotechnology, National Cheng Kung University, Tainan, Taiwan and Institute of Clinical Medicine, College of Medicine, National Cheng Kung University, Tainan, Taiwan.
As immunotoxicity assessments of newly developed biomaterials are often restricted to use in assessment of local tissue response at the implantation site, they do not always show an immune response acceptable to qualify them for clinical use. We tested a new method to assess systemic toxicity: counting the CD4(+) and CD8(+) cells in the spleen. Three different biomaterials were subcutaneously implanted in three groups of rats for the same time period. After 31 days, their spleens were harvested, and CD4(+) and CD8(+) cells were counted. The mean CD4(+)/CD8(+) cell counts were 24.5 ± 3.6/19.8 ± 4.0 (porous collagen matrix group), 25.5 ± 7.1/21.6 ± 3.8 [synthetic collagen matrix (Duragen®) group] and 28.1 ± 4.1/19.6 ± 3.7 (porcine dermis group). Differences in cell counts were not significant. The immunotoxic response generated against porous collagen matrix was comparable to that produced by a similar biomaterial already used clinically. This is, to the best of our knowledge, the first study on cytotoxic lymphocytes in the spleen to quantify systemic immune response to a biomaterial; however, such studies have been conducted with bacterial and viral antigens, and with vaccines. We believe that the present study provides a viable method for larger studies to confirm our current findings.
As immunotoxicity assessments of newly developed biomaterials are often restricted to use in assessment of local tissue response at the implantation site, they do not always show an immune response acceptable to qualify them for clinical use. We tested a new method to assess systemic toxicity: counting the CD4(+) and CD8(+) cells in the spleen. Three different biomaterials were subcutaneously implanted in three groups of rats for the same time period. After 31 days, their spleens were harvested, and CD4(+) and CD8(+) cells were counted. The mean CD4(+)/CD8(+) cell counts were 24.5 ± 3.6/19.8 ± 4.0 (porous collagen matrix group), 25.5 ± 7.1/21.6 ± 3.8 [synthetic collagen matrix (Duragen®) group] and 28.1 ± 4.1/19.6 ± 3.7 (porcine dermis group). Differences in cell counts were not significant. The immunotoxic response generated against porous collagen matrix was comparable to that produced by a similar biomaterial already used clinically. This is, to the best of our knowledge, the first study on cytotoxic lymphocytes in the spleen to quantify systemic immune response to a biomaterial; however, such studies have been conducted with bacterial and viral antigens, and with vaccines. We believe that the present study provides a viable method for larger studies to confirm our current findings.
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