Pia Montanucci

Long-Term Metabolic and Immunological Follow-Up of Nonimmunosuppressed Patients With Type 1 Diabetes Treated With Microencapsulated Islet Allografts Four cases

OBJECTIVE To assess long-term metabolic and immunological follow-up of microencapsulated human islet allografts in nonimmunosuppressed patients with type 1 diabetes (T1DM). RESEARCH DESIGN AND METHODS Four nonimmunosuppressed patients, with long-standing T1DM, received intraperitoneal transplant (TX) of microencapsulated human islets. Anti-major histocompatibility complex (MHC) class I–II, GAD65, and islet cell antibodies were measured before and long term after TX. RESULTS All patients turned positive for serum C-peptide response, both in basal and after stimulation, throughout 3 years of posttransplant follow-up. Daily mean blood glucose, as well as HbA1c levels, significantly improved after TX, with daily exogenous insulin consumption declining in all cases and being discontinued, just transiently, only in patient 4. Anti-MHC class I–II and GAD65 antibodies all tested negative at 3 years after TX. CONCLUSIONS The grafts did not elicit any immune response, even in the cases where more than one preparation was transplanted, as a unique finding, compatible with encapsulation-driven “bioinvisibility” of the grafted islets. This result had never been achieved with the recipient’s general immunosuppression.

In vitro immunomodulatory effects of microencapsulated umbilical cord Wharton jelly-derived mesenchymal stem cells in primary Sjögren’s syndrome

OBJECTIVE Human umbilical cord Wharton jelly-derived mesenchymal stem cells (hUCMS) are easy to retrieve in bulk. They may interact with immune cells by either cell contact or soluble factors. Little evidence is currently available on potential therapeutic application of hUCMS to systemic autoimmune disorders such as primary SS (pSS). We have recently developed an endotoxin-free alginate gel that can be used to microencapsulate different cell types for graft into non-immunosuppressed hosts. We aimed to assess the in vitro effects of IFN-γ-pretreated microencapsulated (CpS)-hUCMS on T cells of pSS. METHODS Ten pSS patients and 10 healthy donors were selected. Peripheral blood mononuclear cells (PBMCs) were obtained from venous blood to establish co-cultures with CpS-hUCMS. Lymphocyte proliferation and phenotypic analysis was performed by flow cytometry and real-time PCR on IFN-γ-pretreated hUCMS was performed before PBMCs co-culture. RESULTS We found that CpS-hUCMS suppress pSS T cell proliferation and restore the Treg/Th17 ratio, thereby possibly positively impacting the pSS disease process. CONCLUSION We have developed a new biohybrid drug delivery system that now waits for clinical application in autoimmune diseases, including pSS.

Stem cells for pancreatic β-cell replacement in diabetes mellitus: actual perspectives.

Purpose of reviewType 1 and type 2 diabetes mellitus represent a widespread metabolic disorder, related to autoimmune &bgr;-cell destruction and insulin resistance, leading to &bgr;-cell dysfunction, respectively, that are associated with severe chronic complications with irreversible multiorgan morphological and functional damage. Conventional treatment, based on exogenous insulin or oral agents may control and delay but not prevent the disease complications, which has lead, so far, to a steady increase in mortality and morbidity. &bgr;-Cell substitution cell therapy, initially pursued by whole pancreatic and isolated islet transplantation, with scarce and limited efficiency, now is looking at the new technologies for cell and molecular therapy for diabetes, based on stem cells. Recent findingsPancreatic endocrine cells regeneration might replenish the destroyed &bgr;-cell pool, with neogenerated &bgr;-cell derived from pancreatic and extrapancreatic stem cell sources. Additionally, embryonic or adult stem cells derived from different cell lineages, and able to differentiate into &bgr;-like cell elements, may not only restore the original insulin secretory patterns but also exert the immunomodulatory effects aimed at interrupting the &bgr;-cell-directed autoimmune destruction vicious cycle. SummaryThese new strategies may, one day, provide for the final cure of diabetes mellitus.

Restoration of t cell substes of patients with type 1 diabetes mellitus by microencapsulated human umbilical cord Wharton jelly-derived mesenchymal stem cells: An in vitro study

Human umbilical cord Wharton jelly-derived mesenchymal stem cells (hUCMS) might apply to treating chronic autoimmune disorders, as already shown for Sjögrens syndrome, including type 1 diabetes mellitus (T1D). Since naked hUCMS grafts encountered restraints, we enveloped hUCMS, within immunoisolatory microcapsules (CpS-hUCMS), made of our endotoxin-free, clinical grade alginate. We then examined the vitro effects of interferon (IFN)-γ-pretreated CpS-hUCMS on Th17 and Treg of T1D patients (n=15) and healthy controls (n=10). Peripheral blood mononuclear cells (PBMCs) were co-cultured with PBMC/CpS-hUCMS: lymphocyte proliferation was assessed by carboxyfluorescein succinimidyl esther (CFSE) dilution assay, and phenotypic analysis of regulatory and effector Tc was also performed. Cytokine expression was performed by bead array and qPCR on IFN-γ-pretreated hUCMS before PBMCs co-culture. CpS-hUCMS restored a correct Treg/Th17 ratio, relevant to the T1D disease process. In summary, we have preliminarily developed a new biohybrid system, associated with immunoregulatory properties, that is ready for in vivo application.

Long-term cultured neonatal porcine islet cell monolayers: a potential tissue source for transplant in diabetes.

Abstract:  Background:  The restricted availability of cadaveric human donor pancreases mandates validation of possibly inexhaustible, alternative sources of insulin secretory cells in order to expand islet transplant for the therapy of insulin dependent diabetes mellitus (T1DM).

Surface Hydrophilicity of Poly(l-Lactide) Acid Polymer Film Changes the Human Adult Adipose Stem Cell Architecture

Current knowledge indicates that the molecular cross-talk between stem cells and biomaterials guides the stem cells’ fate within a tissue engineering system. In this work, we have explored the effects of the interaction between the poly(l-lactide) acid (PLLA) polymer film and human adult adipose stem cells (hASCs), focusing on the events correlating the materials’ surface characteristics and the cells’ plasma membrane. hASCs were seeded on films of pristine PLLA polymer and on a PLLA surface modified by the radiofrequency plasma method under oxygen flow (PLLA+O2). Comparative experiments were performed using human bone-marrow mesenchymal stem cells (hBM-MSCs) and human umbilical matrix stem cells (hUCMSCs). After treatment with oxygen-plasma, the surface of PLLA films became hydrophilic, whereas the bulk properties were not affected. hASCs cultured on pristine PLLA polymer films acquired a spheroid conformation. On the contrary, hASCs seeded on PLLA+O2 film surface maintained the fibroblast-like morphology typically observed on tissue culture polystyrene. This suggests that the surface hydrophilicity is involved in the acquisition of the spheroid conformation. Noteworthy, the oxygen treatment had no effects on hBM-MSC and hUCMSC cultures and both stem cells maintained the same shape observed on PLLA films. This different behavior suggests that the biomaterial-interaction is stem cell specific.

Umbilical cord mesenchymal stem cells for the treatment of autoimmune diseases: beware of cell-to-cell contact

We have read with great interest the article from Deng and colleagues1 reporting on the results of a randomised, controlled clinical trial of allogeneic umbilical cord-derived mesenchymal stem cells (UC-MSCs) for lupus nephritis. The study demonstrated that the administration of intravenous UC-MSCs does not provide additional benefit to conventional immunosuppression. Over the last decade, increasing evidence has indicated that UC-MSCs are multipotent cells with promising applications to regenerative medicine2 and also a powerful tool to modulate the immune system. UC-MSCs can interact with most immune cell types via both cell–cell contact and secretion …

Microencapsulation of Islets for the Treatment of Type 1 Diabetes Mellitus (T1D)

Microencapsulation technology, based on use of alginic acid biopolymers, has been devised many years ago. However, when intended for enveloping human islets for transplantation purposes, the method needs to be up-scaled and implemented with care being taken to comply with simple but important measures. It is almost indispensable to rely on an ultrapurified alginic polymers: in fact, any, even minimal, alginate contamination with endotoxins, pyrogens, and proteins could provoke the hosts inflammatory reaction upon graft, with heavy adverse consequences on the capsules immunoprotective properties, hence on graft survival per se. Care should be taken in ensuring fabrication of reproducible microspheres, in terms not only of shape and size, but also consistency of the peripheral layers around the central alginate gel core, where the islets are immobilized. Once the product is well defined and stable, care should also be taken in accurately selecting patients with T1D that are candidate for encapsulated islet cell transplantation with no general immunosuppression. A series of pre- and post-intraperitoneal transplant metabolic, chemical, and immunological parameters are to be monitored, in conjunction with image analysis of the abdomen, in order to assess efficacy of the intervention according to well defined grading scale.

A Comparison of Lysosomal Enzymes Expression Levels in Peripheral Blood of Mild- and Severe-Alzheimer’s Disease and MCI Patients: Implications for Regenerative Medicine Approaches

The association of lysosomal dysfunction and neurodegeneration has been documented in several neurodegenerative diseases, including Alzheimer’s Disease (AD). Herein, we investigate the association of lysosomal enzymes with AD at different stages of progression of the disease (mild and severe) or with mild cognitive impairment (MCI). We conducted a screening of two classes of lysosomal enzymes: glycohydrolases (β-Hexosaminidase, β-Galctosidase, β-Galactosylcerebrosidase, β-Glucuronidase) and proteases (Cathepsins S, D, B, L) in peripheral blood samples (blood plasma and PBMCs) from mild AD, severe AD, MCI and healthy control subjects. We confirmed the lysosomal dysfunction in severe AD patients and added new findings enhancing the association of abnormal levels of specific lysosomal enzymes with the mild AD or severe AD, and highlighting the difference of AD from MCI. Herein, we showed for the first time the specific alteration of β-Galctosidase (Gal), β-Galactosylcerebrosidase (GALC) in MCI patients. It is notable that in above peripheral biological samples the lysosomes are more sensitive to AD cellular metabolic alteration when compared to levels of Aβ-peptide or Tau proteins, similar in both AD groups analyzed. Collectively, our findings support the role of lysosomal enzymes as potential peripheral molecules that vary with the progression of AD, and make them useful for monitoring regenerative medicine approaches for AD.

Functional Profiles of Human Umbilical Cord-Derived Adult Mesenchymal Stem Cells in Obese/Diabetic Versus Healthy Women

BACKGROUND Adult human mesenchymal stem cells retrieved, from the post-partum human umbilical cord Wharton jelly (hUCMS), have recently gained growing interest due to their morphological and functional properties. OBJECTIVE The main purpose of our work was to examine morphology and functional properties of hUCMS retrieved from healthy women as compared to those with obesity, or gestational or type 2 diabetes mellitus, under fair metabolic control. Possible differences between groups could shed light into the potential use of these cells for the cell therapy of a variety of diseases, regardless of the obesity/diabetes status of the donor mothers. Additionally, information on how the maternal disease may affect the cord-derived stem cells, hence possibly newborn children would be important. METHOD We have studied obese/diabetic or normal donor post-partum umbilical cord-derived hUCMS, either in basal or during differentiation protocols into several cell phenotypes and the definitive endoderm. Immunomodulatory properties of these cells, in terms of inhibition of activated lymphocyte proliferation, also was examined. RESULTS According to our preliminary results, there are functional differences, as assessed by cell and molecular assays, in terms of both, differentiation and immunomodulatory potential, between the cells derived from normal as compared to obese/diabetic mothers. CONCLUSION The findings seemingly indicate that the uterine environment of obese/diabetic mothers is quite distant from normal, regardless of metabolic control. Hence hUCMS extracted from obese/diabetic mothers do not appear to be suitable for cell therapy clinical protocols but more studies are required.