Journal: Bioactive Materials

Article Title: Cryopreservable, scalable and ready-to-use cell-laden patches for diabetic ulcer treatment

doi: 10.1016/j.bioactmat.2025.04.024

Figure Lengend Snippet: Fabrication, cryopreservation and structure maintenance of ADSC-patch. ( A ) UV–vis spectrum of synthesized Alg-DA. ( B ) Adhesion of ADSC-patch to outstretched finger and digital joint. Methylthioninium chloride labeled ADSC-patch before cryopreservation, rhodamine labeled ADSC-patch after cryopreservation. ( C ) ADSCs viability in Alg-DA hydrogel patches on Day 7. ADSC-patch and Cryo ADSC-patch refers to fresh ADSC-patch and cryopreserved ADSC-patch, respectively. ( D ) Tunel staining of ADSCs in normal culture and ADSC-patches to characterize apoptosis. ( E ) SEM images of ADSC-patches before and after cryopreservation. Comparisons of cryopreservation pathways between this study and other conventional approaches in ( F ) phase diagram, ( G ) thermal time course, and ( H ) cytotoxic (penetrating CPA, p CPA) time course. ld and ud represent CPA loading and unloading steps before and after cryopreservation, respectively. Arrows indicate the direction of the preservation process. The axes are not drawn to scale. The rates of temperature and osmolality change are represented by the slopes of the curves. Long-term storage is denoted by ‘//’. ( I ) Storage modulus of various hydrogel patches before and after cryopreservation. ( J ) Statistics on structural integrity of 0.5 mm, 0.75 mm and 1 mm thick hydrogel patches after fabrication. ( K ) Statistics on structural integrity of 0.5 mm, 0.75 mm and 1 mm thick hydrogel patches after cryopreservation in −86 °C and in LN 2 (−196 °C). ( L ) Representative images of 0.5 mm, 0.75 mm and 1 mm thick hydrogel patches after cryopreservation in LN 2 (−196 °C). 10 DMSO and 5D+5P represent cryopreservation with 10 % DMSO and 5 % DMSO+5 % PROH, respectively. The dashed line represents the broken ADSC-patch, while the solid line represents the intact ADSC-patch Independent repeat experiment n ≥ 3. ∗: p < 0.05, ∗∗: p < 0.01, and ∗∗∗: p < 0.001.

Article Snippet: Next, SD rat ADSCs (P4-P6, Cyagen) were resuspended in 2 % (w/v) hydrogel precursor solution (Alg or Alg-DA dissolved in DMEM) and the resulting mixture was added dropwise into PDMS molds (87.5 μL/per hole), then the molds were covered uniformly with dialysis membrane, and finally an appropriate amount of 100 mM CaCl 2 solution was added dropwise on top of the dialysis membrane.

Techniques: Synthesized, Labeling, TUNEL Assay, Staining, Preserving

Journal: Bioactive Materials

Article Title: Cryopreservable, scalable and ready-to-use cell-laden patches for diabetic ulcer treatment

doi: 10.1016/j.bioactmat.2025.04.024

Figure Lengend Snippet: Treatment of diabetic ulcers by cryopreserved ADSC-patch. ( A ) In vivo bioluminescence imaging, and ( B ) quantitative analysis of the Luc + /GFP + ADSCs delivered by ADSC suspension, fresh and cryopreserved ADSC-patches. n = 4. ( C ) Photographs of diabetic ulcers healing of 6 groups: 1) untreated (Control); 2) hydrogel alone (Alg-DA); 3) ADSCs alone (ADSCs); 4) fresh ADSC-patch (ADSC-patch); 5) cryopreserved ADSC-patch (Cryo ADSC-patch); 6) cryopreserved ADSC-patch with 24-h recovery (Cryo/rec ADSC-patch). n = 18. ( D ) Quantitative analysis of wound size normalized to Day 0. ( E ) Quantitative analysis of scar area on Day 14. ( F ) Representative H&E stained images of diabetic ulcers on Day 14. Blue dotted line represents the unclosed area of the ulcer. Solid and open arrows respectively point to granulation and re-epithelialization in regenerated tissues. Independent repeat experiment n ≥ 3. ∗: p < 0.05, ∗∗: p < 0.01, and ∗∗∗: p < 0.001.

Article Snippet: Next, SD rat ADSCs (P4-P6, Cyagen) were resuspended in 2 % (w/v) hydrogel precursor solution (Alg or Alg-DA dissolved in DMEM) and the resulting mixture was added dropwise into PDMS molds (87.5 μL/per hole), then the molds were covered uniformly with dialysis membrane, and finally an appropriate amount of 100 mM CaCl 2 solution was added dropwise on top of the dialysis membrane.

Techniques: In Vivo, Imaging, Suspension, Control, Staining