Shariati, K. | Search bioRxiv

bioRxiv2025

Resorbable Poly-(D,L)-Lactide Anchorage of Nanoparticulate Mineralized Collagen Materials Maximizes In Vivo Skull Regeneration

BackgroundA clinical demand exists for bone biomaterials that mirror tissue-specific extracellular matrix (ECM) properties and regulate progenitor cell fate. Nanoparticulate mineralized collagen glycosaminoglycan (MC-GAG) scaffolds have demonstrated to promote skull regeneration in vivo without addition of growth factors or exogenous progenitor cells, offering a materials-only solution for cranial defect reconstructions. Further enhancement of the safety and regenerative potential of MC-GAG is however necessary for clinical translation. Here, we investigated the regenerative effect of MC-GAG scaffold fixation with a resorbable poly(D,L-lactide) (PDLLA) implant for cerebral protection. MethodsFourteen-millimeter cranial defects were created in New Zealand white rabbits, divided into four groups: 1) defect only, 2) defect with PDLLA, 3) MC-GAG, and 4) MC-GAG with PDLLA. Initial bone healing assessment was conducted at 3 and 9 months using microcomputed tomography (microCT), histology, reference point indentation, and scanning electron microscopy. Long-term effects were evaluated through in vivo microCT imaging at 3, 6, and 9 months, with biomechanical testing of explanted skulls at 9 months. ResultsAt 3 months, MC-GAG significantly enhanced mineralization compared to empty and PDLLA-treated defects, with even greater mineralization observed for MC-GAG-PDLLA. Histologically, MC-GAG demonstrated trabecular mineralization, while MC-GAG-PDLLA had a structural organization resembling native bone. Biomechanical assessment showed enhanced toughness and stiffness for both MC-GAG and MC-GAG-PDLLA. At 6 months, MC-GAG-PDLLA displayed the highest bone mass, and at 9 months, both MC-GAG-containing scaffolds demonstrated superior bone regeneration compared to controls. By 9 months, PDLLA implants were completely resorbed and MC-GAG-PDLLA exhibited mechanical properties surpassing those of all other groups. ConclusionsFixation of MC-GAG scaffolds with PDLLA implants improved mineralization and bone regeneration compared to either material alone for cranial defect reconstruction. Clinical Relevance StatementCranioplasties are essential for cerebral protection and neurological restoration, but current materials have limitations, necessitating alternative bone biomaterials for improved outcomes.

bioengineering↗

bioRxiv2024

An shRNA screen in primary human beta cells identifies the serotonin 1F receptor as a negative regulator of survival during transplant

Islet transplantation can cure type 1 diabetes, but peri-transplant beta cell death limits this procedure to those with low insulin requirements. Improving human beta cell survival or proliferation may make islet transplantation a possibility for more type 1 patients. To identify novel regulators of beta cell survival and proliferation, we conducted a pooled small hairpin RNA (shRNA) screen in primary human beta cells transplanted into immunocompromised mice. shRNAs targeting several cyclin dependent kinase inhibitors were enriched after transplant. Here, we focused on the Gi/o-coupled GPCR, serotonin 1F receptor (HTR1F, 5-HT1F) which our screen identified as a negative regulator of beta cell numbers after transplant. In vitro, 5-HT1F knockdown induced human beta cell proliferation but only when combined with harmine and exendin-4. In vivo, knockdown of 5-HT1F reduced beta cell death during transplant. To demonstrate the feasibility of targeting 5-HT1F in islet transplant, we identified and validated a small molecule 5-HT1F antagonist. This antagonist increased glucose stimulated insulin secretion from primary human islets and cAMP accumulation in primary human beta cells. Finally, the 5-HT1F antagonist improved glycemia in marginal mass, human islet transplants into immunocompromised mice. We identify 5-HT1F as a novel druggable target to improve human beta cell survival in the setting of islet transplantation. One Sentence SummarySerotonin 1F receptor (5-HT1F) negatively regulates insulin secretion and beta cell survival during transplant.

cell biology↗