Journal: Nature Communications

Article Title: Implantable niche with local immunosuppression for islet allotransplantation achieves type 1 diabetes reversal in rats

doi: 10.1038/s41467-022-35629-z

Figure Lengend Snippet: a Optical image of NICHE and annotated rendering of NICHE and scanning electron microscopy (SEM) images of the two-layer mesh and nanoporous membrane (representative scan images of 5 independent samples). The longitudinal section of NICHE is representative for clarity. Representative Masson’s trichrome staining of fibrotic capsule around b resin and c PA devices implanted in rats for 6 weeks. d Quantification of fibrotic capsule thickness around medical grade titanium (Ti; n = 5), resin ( n = 5), and PA ( n = 4) devices implanted subQ for 6 weeks in rats, mean ± SD, one-way analysis of variance (ANOVA) *** p < 0.001. e Implant r e activity scores of Ti ( n = 5), resi n ( n = 5), and PA ( n = 4) devices implanted subQ for 6 weeks in rats, mean ± SD, one-way ANOVA, n.s. p = 0.1117. Optical images showing cross sections of f resin and g PA NICHE implanted subQ for 6 weeks. Dashed lines indicate cell reservoir. h Resin and PA roughness quantified using atomic force microscopy ( n = 9 devices per material), mean ± SD, unpaired two-tailed student’s t-test ** p < 0.01. i Contact angle analysis of resin and PA devices ( n = 3 devices per material), mean ± SD, unpaired two-tailed student’s t-test *** p < 0.001. j Resin and PA NICHE cell reservoir tissue sections stained with blood vessel marker B. simplicifolia lectin; vessels in red pointed by black arrow heads. k Vascular density quantification of resin ( n = 1) and PA ( n = 4) cell reservoir tissue from independent NICHE, mean ± SD. l Daily percent in vitro release of CTLA4Ig and IgG from NICHE drug reservoir ( n = 4 devices per drug), mean ± SD. Static glucose stimulated insulin release (GSIR) of rat islets after 5 days of incubation with m CTLA4Ig, n ALS, and o CTLA4Ig + ALS ( n = 4 per condition), mean ± SD, o n e-way ANOVA, * p < 0.05, ** p < 0.001, n.s. p > 0.05 of each condition versus vehicle (0 mg/mL). Source data are provided as a Source Data file.

Article Snippet: The immunosuppressants used for rat studies were CTLA4Ig (Abatacept, Bristol-Myers Squibb) and rabbit anti-lymphocyte serum (ALS; Accurate chemical); and for NHP studies anti-thymocyte globulin (ATG; Sanofi), CTLA4Ig (Belatacept, Bristol-Myers Squibb), and anti-CD154 [5C8H1] (NIH Nonhuman Primate Reagent Resource, RRID: AB_2716324).

Techniques: Electron Microscopy, Membrane, Staining, Activity Assay, Microscopy, Two Tailed Test, Marker, In Vitro, Incubation

Journal: Nature Communications

Article Title: Implantable niche with local immunosuppression for islet allotransplantation achieves type 1 diabetes reversal in rats

doi: 10.1038/s41467-022-35629-z

Figure Lengend Snippet: a In vitro tube formation activity of human umbilical vein endothelial cells (HUVEC) in the presence of different drug concentrations (mg/mL). Supplemented media was used as positive inducer (+), 30 µM suramin as positive inhibitor (-), and non-supplemented media as control (Veh). b Total length quantification and c number of segments of tube formation ( n = 4 biological replicates per condition), mean ± SD, one-way ANOVA with Tukey’s multiple comparisons test (*** p < 0.001). d qPCR analysis of vascular related genes in HUVEC after treatment with different concentrations of ATG and CTLA4Ig (mg/mL). Gapdh gene was used as internal control and fold-change in gene expression is relative to control untreated cells. ( n = 4 biological replicates), mean ± SD, one-way ANOVA with Tukey’s multiple comparisons test (*** p < 0.001; n.s. p > 0.05). e Quantification of VEGF levels in cell reservoir and surrounding microenvironment for control and IS release samples ( n = 4 biological replicates), mea n ± SD, unpaired two-tailed student’s t -test n.s. p > 0.05. f Fluorescence intensity measurements of IHC analysis in g ( n = 4 biological replicates), mean ± SD, unpaired two-tailed student’s t -test n.s. p > 0.05. g Representative sections of NICHE control and IS release slides stained with functional blood vessel markers CD31 (red), VE-cadherin (green), and eNOS (gold). Source data are provided as a Source Data file.

Article Snippet: The immunosuppressants used for rat studies were CTLA4Ig (Abatacept, Bristol-Myers Squibb) and rabbit anti-lymphocyte serum (ALS; Accurate chemical); and for NHP studies anti-thymocyte globulin (ATG; Sanofi), CTLA4Ig (Belatacept, Bristol-Myers Squibb), and anti-CD154 [5C8H1] (NIH Nonhuman Primate Reagent Resource, RRID: AB_2716324).

Techniques: In Vitro, Activity Assay, Control, Gene Expression, Two Tailed Test, Fluorescence, Staining, Functional Assay

Journal: Nature Communications

Article Title: Implantable niche with local immunosuppression for islet allotransplantation achieves type 1 diabetes reversal in rats

doi: 10.1038/s41467-022-35629-z

Figure Lengend Snippet: Plasma levels of a ALS and b CTLA4Ig of NICHE ( n = 12 to day 63, n = 8 to day 92, n = 5 to day 112, n = 3 to day 192) and IP ( n = 8 to day 63, n = 5 to day 112) rats, mean ± SD, unpaired two-tailed student’s t -test between NICHE and IP overall average concentrations, (*** p < 0.001). Black triangles indicate drug reservoir reloading. The red triangle indicates systemic ALS bolus. LOQ limit of quantitation. Quantification of immunosuppressants in c peripheral organs and d transplant microenvironment at endpoint of NICHE and IP rats ( n = 8), mea n ± SD, unpaired two-tailed student’s t -test between NICHE and IP rats for each organ (* p < 0.05; ** p < 0.01; *** p < 0.001; n.s. p > 0.05; n.q. = not quantifiable). FC = fibrotic capsule. e Quantification of lymphocytes in blood expressed as fold change to day 0 of NICHE ( n = 8 to day 56, n = 5 to day 115), IP ( n = 5), and rats receiving vehicle ( n = 4). f Quantification of Tregs in blood on day 84 of NICHE, IP, and healthy rats ( n = 5), mean ± SD, one-way ANOVA with Tukey’s multiple comparisons test (*** p < 0.001; n.s. p = 0.1643). Source data are provided as a Source Data file.

Article Snippet: The immunosuppressants used for rat studies were CTLA4Ig (Abatacept, Bristol-Myers Squibb) and rabbit anti-lymphocyte serum (ALS; Accurate chemical); and for NHP studies anti-thymocyte globulin (ATG; Sanofi), CTLA4Ig (Belatacept, Bristol-Myers Squibb), and anti-CD154 [5C8H1] (NIH Nonhuman Primate Reagent Resource, RRID: AB_2716324).

Techniques: Clinical Proteomics, Two Tailed Test, Quantitation Assay