Association between Mortality from Chronic Noncommunicable Diseases and Human Development Index in Brazil between 1980 and 2019. / Associação entre a Mortalidade por Doenças Crônicas Não Transmissíveis e o Índice de Desenvolvimento Humano no Brasil entre 1980 e 2019.

BACKGROUND: Chronic noncommunicable diseases (CNCDs) caused more than 734,000 deaths (55% of all deaths) in Brazil in 2019, with an important socioeconomic impact. OBJECTIVES: To analyze the mortality rates from CNCDs in Brazil from 1980 to 2019 and their association with socioeconomic indicators. METHOD: This was a descriptive, time-series study of deaths from CNCDs in Brazil from 1980 to 2019. Data on the annual frequencies of deaths and on population were obtained from the Department of Informatics of the Brazilian Unified Health System. Crude and standardized mortality rates per 100,000 inhabitants were estimated using the direct method (Brazilian population in 2000). The quartiles of each CNCD were calculated, where a quartile change, due to an increase in mortality rate, was represented by a chromatic gradient. The Municipal Human Development Index (MHDI) of each Brazilian federative unit was extracted from the Atlas Brasil website and correlated with the rates of CNCD mortality. RESULTS: There was a reduction in mortality rates due to diseases of the circulatory system during the period, except in the Northeast Region. There was also an increase in mortality from neoplasia and diabetes, while the rates of chronic respiratory diseases showed little variation. There was an inverse correlation between the federative units with greater reduction in CNCD mortality rates and the MHDI. CONCLUSIONS: The observed decrease in mortality due to diseases of the circulatory system may reflect an improvement in socioeconomic indicators in Brazil during the period. The increase in mortality rates due to neoplasms is probably related to the aging of the population. The higher mortality rates of diabetes seem to be associated with an increase in the prevalence of obesity in Brazilian women.

FUNDAMENTO: No Brasil, em 2019, as doenças crônicas não transmissíveis (DCNT) acarretaram mais de 734 mil óbitos, 55% de todas as mortes, com importante impacto socioeconômico. OBJETIVOS: Analisar as taxas de mortalidade das DCNT, no Brasil, de 1980 a 2019, e sua associação com indicadores socioeconômicos. MÉTODOS: Trata-se de um estudo descritivo, de séries temporais dos óbitos por DCNT, no Brasil, de 1980 a 2019. Os dados relativos às frequências anuais de mortes e da população foram obtidos do DATASUS. Foram estimadas as taxas de mortalidade brutas e padronizadas por 100.000 habitantes, pelo método direto (população do Brasil de 2000). Foram calculados os quartis de cada DCNT, onde a mudança de quartil, por aumento das taxas de mortalidade, foi representada por gradiente cromático. O Índice de Desenvolvimento Humano Municipal (IDHM) de cada unidade da federação (UF) foi extraído do site Atlas Brasil e correlacionado com as taxas de mortalidade por DCNT. RESULTADOS: Ocorreu redução nas taxas de mortalidade por doenças do aparelho circulatório no período, exceto na região Nordeste. Houve também aumento da mortalidade por neoplasia e diabetes, enquanto as taxas das doenças respiratórias apresentaram poucas variações. Houve correlação inversa entre as UF com maior redução nas taxas de mortalidade por DCNT e o IDHM. CONCLUSÕES: A redução observada na mortalidade por doenças do aparelho circulatório pode refletir melhoria dos indicadores socioeconômicos, no Brasil, nesse período. O aumento da taxa de mortalidade por neoplasias provavelmente se relaciona com o envelhecimento da população. As maiores taxas de mortalidade por diabetes parecem ser associadas com o aumento da prevalência da obesidade nas mulheres brasileiras.

Good Manufacturing Practice-compliant change of raw material in the manufacturing process of a clinically used advanced therapy medicinal product-a comparability study.

The development of medicinal products often continues throughout the different phases of a clinical study and may require challenging changes in raw and starting materials at later stages. Comparability between the product properties pre- and post-change thus needs to be ensured. Here, we describe and validate the regulatory compliant change of a raw material using the example of a nasal chondrocyte tissue-engineered cartilage (N-TEC) product, initially developed for treatment of confined knee cartilage lesions. Scaling up the size of N-TEC as required for the treatment of larger osteoarthritis defects required the substitution of autologous serum with a clinical-grade human platelet lysate (hPL) to achieve greater cell numbers necessary for the manufacturing of larger size grafts. A risk-based approach was performed to fulfill regulatory requirements and demonstrate comparability of the products manufactured with the standard process (autologous serum) already applied in clinical indications and the modified process (hPL). Critical attributes with regard to quality, purity, efficacy, safety and stability of the product as well as associated test methods and acceptance criteria were defined. Results showed that hPL added during the expansion phase of nasal chondrocytes enhances proliferation rate, population doublings and cell numbers at passage 2 without promoting the overgrowth of potentially contaminant perichondrial cells. N-TEC generated with the modified versus standard process contained similar content of DNA and cartilaginous matrix proteins with even greater expression levels of chondrogenic genes. The increased risk for tumorigenicity potentially associated with the use of hPL was assessed through karyotyping of chondrocytes at passage 4, revealing no chromosomal changes. Moreover, the shelf-life of N-TEC established for the standard process could be confirmed with the modified process. In conclusion, we demonstrated the introduction of hPL in the manufacturing process of a tissue engineered product, already used in a late-stage clinical trial. Based on this study, the national competent authorities in Switzerland and Germany accepted the modified process which is now applied for ongoing clinical tests of N-TEC. The described activities can thus be taken as a paradigm for successful and regulatory compliant demonstration of comparability in advanced therapy medicinal products manufacturing.

Associação entre a Mortalidade por Doenças Crônicas Não Transmissíveis e o Índice de Desenvolvimento Humano no Brasil entre 1980 e 2019 / Association between Mortality from Chronic Noncommunicable Diseases and Human Development Index in Brazil between 1980 and 2019

Resumo Fundamento No Brasil, em 2019, as doenças crônicas não transmissíveis (DCNT) acarretaram mais de 734 mil óbitos, 55% de todas as mortes, com importante impacto socioeconômico. Objetivos Analisar as taxas de mortalidade das DCNT, no Brasil, de 1980 a 2019, e sua associação com indicadores socioeconômicos. Métodos Trata-se de um estudo descritivo, de séries temporais dos óbitos por DCNT, no Brasil, de 1980 a 2019. Os dados relativos às frequências anuais de mortes e da população foram obtidos do DATASUS. Foram estimadas as taxas de mortalidade brutas e padronizadas por 100.000 habitantes, pelo método direto (população do Brasil de 2000). Foram calculados os quartis de cada DCNT, onde a mudança de quartil, por aumento das taxas de mortalidade, foi representada por gradiente cromático. O Índice de Desenvolvimento Humano Municipal (IDHM) de cada unidade da federação (UF) foi extraído do site Atlas Brasil e correlacionado com as taxas de mortalidade por DCNT. Resultados Ocorreu redução nas taxas de mortalidade por doenças do aparelho circulatório no período, exceto na região Nordeste. Houve também aumento da mortalidade por neoplasia e diabetes, enquanto as taxas das doenças respiratórias apresentaram poucas variações. Houve correlação inversa entre as UF com maior redução nas taxas de mortalidade por DCNT e o IDHM. Conclusões A redução observada na mortalidade por doenças do aparelho circulatório pode refletir melhoria dos indicadores socioeconômicos, no Brasil, nesse período. O aumento da taxa de mortalidade por neoplasias provavelmente se relaciona com o envelhecimento da população. As maiores taxas de mortalidade por diabetes parecem ser associadas com o aumento da prevalência da obesidade nas mulheres brasileiras.

Abstract Background Chronic noncommunicable diseases (CNCDs) caused more than 734,000 deaths (55% of all deaths) in Brazil in 2019, with an important socioeconomic impact. Objectives To analyze the mortality rates from CNCDs in Brazil from 1980 to 2019 and their association with socioeconomic indicators. Method This was a descriptive, time-series study of deaths from CNCDs in Brazil from 1980 to 2019. Data on the annual frequencies of deaths and on population were obtained from the Department of Informatics of the Brazilian Unified Health System. Crude and standardized mortality rates per 100,000 inhabitants were estimated using the direct method (Brazilian population in 2000). The quartiles of each CNCD were calculated, where a quartile change, due to an increase in mortality rate, was represented by a chromatic gradient. The Municipal Human Development Index (MHDI) of each Brazilian federative unit was extracted from the Atlas Brasil website and correlated with the rates of CNCD mortality. Results There was a reduction in mortality rates due to diseases of the circulatory system during the period, except in the Northeast Region. There was also an increase in mortality from neoplasia and diabetes, while the rates of chronic respiratory diseases showed little variation. There was an inverse correlation between the federative units with greater reduction in CNCD mortality rates and the MHDI. Conclusions The observed decrease in mortality due to diseases of the circulatory system may reflect an improvement in socioeconomic indicators in Brazil during the period. The increase in mortality rates due to neoplasms is probably related to the aging of the population. The higher mortality rates of diabetes seem to be associated with an increase in the prevalence of obesity in Brazilian women.

Comparison of Human Articular Cartilage Tissue and Chondrocytes Isolated from Peripheral versus Central Regions of Traumatic Lesions.

OBJECTIVE: Cellular and molecular events occurring in cartilage regions close to injury are poorly investigated, but can possibly compromise the outcome of cell-based cartilage repair. In this study, key functional properties were assessed for cartilage biopsies collected from the central part of traumatic joint lesions (central) and from regions surrounding the defect (peripheral). These properties were then correlated with the quality of the initial cartilage biopsy and the inflammatory state of the joint. DESIGN: Cartilage samples were collected from knee joints of 42 patients with traumatic knee injuries and analyzed for cell phenotype (by reverse transcriptas-polymerase chain reaction), histological quality, cellularity, cell viability, proliferation capacity, and post-expansion chondrogenic capacity of chondrocytes (in pellet culture). Synovium was also harvested and analyzed for the expression of inflammatory cytokines. RESULTS: Cartilage quality and post-expansion chondrogenic capacity were higher in peripheral versus central samples. Differences between these 2 parameters were more pronounced in joints with high inflammatory features characterized by >100-fold difference in the mRNA levels of IL6 and IL8 in the corresponding synovium. Peripheral chondrocytes isolated from good- versus bad-quality biopsies expressed higher levels of collagen II/I and aggrecan/versican and lower levels of MMP13 and ADAMTS5. They also exhibited reduced proliferation and enhanced cartilage-forming capacity. CONCLUSIONS: Chondrocytes at the periphery of traumatic lesions better maintain properties of healthy cartilage compared to those isolated from the center, even when derived from bad-quality tissues harvested from highly inflamed joints. Future studies are necessary to investigate the change of functional properties of peripheral chondrocytes over time.

Nasal chondrocyte-based engineered autologous cartilage tissue for repair of articular cartilage defects: an observational first-in-human trial.

BACKGROUND: Articular cartilage injuries have poor repair capacity, leading to progressive joint damage, and cannot be restored predictably by either conventional treatments or advanced therapies based on implantation of articular chondrocytes. Compared with articular chondrocytes, chondrocytes derived from the nasal septum have superior and more reproducible capacity to generate hyaline-like cartilage tissues, with the plasticity to adapt to a joint environment. We aimed to assess whether engineered autologous nasal chondrocyte-based cartilage grafts allow safe and functional restoration of knee cartilage defects. METHODS: In a first-in-human trial, ten patients with symptomatic, post-traumatic, full-thickness cartilage lesions (2-6 cm2) on the femoral condyle or trochlea were treated at University Hospital Basel in Switzerland. Chondrocytes isolated from a 6 mm nasal septum biopsy specimen were expanded and cultured onto collagen membranes to engineer cartilage grafts (30 × 40 × 2 mm). The engineered tissues were implanted into the femoral defects via mini-arthrotomy and assessed up to 24 months after surgery. Primary outcomes were feasibility and safety of the procedure. Secondary outcomes included self-assessed clinical scores and MRI-based estimation of morphological and compositional quality of the repair tissue. This study is registered with ClinicalTrials.gov, number NCT01605201. The study is ongoing, with an approved extension to 25 patients. FINDINGS: For every patient, it was feasible to manufacture cartilaginous grafts with nasal chondrocytes embedded in an extracellular matrix rich in glycosaminoglycan and type II collagen. Engineered tissues were stable through handling with forceps and could be secured in the injured joints. No adverse reactions were recorded and self-assessed clinical scores for pain, knee function, and quality of life were improved significantly from before surgery to 24 months after surgery. Radiological assessments indicated variable degrees of defect filling and development of repair tissue approaching the composition of native cartilage. INTERPRETATION: Hyaline-like cartilage tissues, engineered from autologous nasal chondrocytes, can be used clinically for repair of articular cartilage defects in the knee. Future studies are warranted to assess efficacy in large controlled trials and to investigate an extension of indications to early degenerative states or to other joints. FUNDING: Deutsche Arthrose-Hilfe.

Regenerative Potential of Tissue-Engineered Nasal Chondrocytes in Goat Articular Cartilage Defects.

Nasal chondrocytes (NC) were previously demonstrated to remain viable and to participate in the repair of articular cartilage defects in goats. Here, we investigated critical features of tissue-engineered grafts generated by NC in this large animal model, namely cell retention at the implantation site, architecture and integration with adjacent tissues, and effects on subchondral bone changes. In this study, isolated autologous goat NC (gNC) and goat articular chondrocytes (gAC, as control) were expanded, green fluorescent protein-labelled and seeded on a type I/III collagen membrane. After chondrogenic differentiation, tissue-engineered grafts were implanted into chondral defects (6 mm in diameter) in the stifle joint for 3 or 6 months. At the time of explantation, surrounding tissues showed no or very low (only in the infrapatellar fat pad <0.32%) migration of the grafted cells. In repair tissue, gNC formed typical structures of articular cartilage, such as flattened cells at the surface and column-like clusters in the middle layers. Semi-quantitative histological evaluation revealed efficient integration of the grafted tissues with the adjacent native cartilage and underlying subchondral bone. A significantly increased subchondral bone area, as a sign for the onset of osteoarthritis, was observed following treatment of cartilage defects with gAC-, but not with gNC-grafts. Our results reinforce the use of NC-based engineered tissue for articular cartilage repair and preliminarily indicate their potential for the treatment of early osteoarthritic defects.

Notochordal cell conditioned medium (NCCM) regenerates end-stage human osteoarthritic articular chondrocytes and promotes a healthy phenotype.

BACKGROUND: Notochordal cell conditioned medium (NCCM) derived from non-chondrodystrophic dogs has pro-anabolic and anti-catabolic effects upon nucleus pulposus (NP) cells. Here, for the first time, we assessed the ability of NCCM to influence the production of extracellular matrix and inflammatory proteins by healthy and osteoarthritic human chondrocytes within engineered cartilage tissues. We hypothesized that, similar to its action on NP cells, NCCM exerts metabolic and anti-catabolic effects on human articular chondrocytes and has the potential to significantly counteract inflammatory mediators. METHODS: Chondrocytes from nine non-osteoarthritic patients and from six osteoarthritic (OA) donors at the time of total knee arthroplasty were chondro-differentiated in pellets for 2 weeks. Non-OA pellets were exposed for 72 hours to IL-1ß/TNF-α and then cultured up to 14 days in 2 % FBS-supplemented NCCM or 2 % FBS-supplemented medium (control (ctr)). OA pellets were cultured in NCCM or ctr medium without pro-inflammatory treatment. Tissues after each culture phase were analyzed biochemically (GAG/DNA), (immuno-) histologically (collagen I, II and GAG) and by Western blotting. Supernatants were analyzed by ELISA. RESULTS: Response to NCCM was age and disease dependent with healthy chondrocyte pellets (from donors >55 years of age) recovering their glycosaminoglycan (GAG) contents to baseline levels only with NCCM. OA pellets treated with NCCM significantly increased GAG content (1.8-fold) and levels of hyaluronic acid link protein (HAPLN), fibromodulin and SOX-9. The catabolic proteins (matrix metalloproteinase (MMP)-3 and MMP-13) and pro-inflammatory enzyme levels (cyclooxygenase-2 (COX-2)) were markedly reduced and there was significantly reduced secretion of pro-inflammatory chemokines (IL-6 and IL-8). CONCLUSIONS: NCCM restores cartilage matrix production of end-stage human OA chondrocytes towards a healthy phenotype and suppresses the production of inflammatory mediators. Harnessing the necessary and sufficient factors within NCCM that confers chondroprotection and regenerative effects could lead to a minimally invasive agent for treatment of degenerative and inflammatory joint diseases.

Novel Perfused Compression Bioreactor System as an in vitro Model to Investigate Fracture Healing.

Secondary bone fracture healing is a physiological process that leads to functional tissue regeneration via endochondral bone formation. In vivo studies have demonstrated that early mobilization and the application of mechanical loads enhances the process of fracture healing. However, the influence of specific mechanical stimuli and particular effects during specific phases of fracture healing remain to be elucidated. In this work, we have developed and provided proof-of-concept of an in vitro human organotypic model of physiological loading of a cartilage callus, based on a novel perfused compression bioreactor (PCB) system. We then used the fracture callus model to investigate the regulatory role of dynamic mechanical loading. Our findings provide a proof-of-principle that dynamic mechanical loading applied by the PCB can enhance the maturation process of mesenchymal stromal cells toward late hypertrophic chondrocytes and the mineralization of the deposited extracellular matrix. The PCB provides a promising tool to study fracture healing and for the in vitro assessment of alternative fracture treatments based on engineered tissue grafts or pharmaceutical compounds, allowing for the reduction of animal experiments.

Bone-forming capacity of adult human nasal chondrocytes.

Nasal chondrocytes (NC) derive from the same multipotent embryological segment that gives rise to the majority of the maxillofacial bone and have been reported to differentiate into osteoblast-like cells in vitro. In this study, we assessed the capacity of adult human NC, appropriately primed towards hypertrophic or osteoblastic differentiation, to form bone tissue in vivo. Hypertrophic induction of NC-based micromass pellets formed mineralized cartilaginous tissues rich in type X collagen, but upon implantation into subcutaneous pockets of nude mice remained avascular and reverted to stable hyaline-cartilage. In the same ectopic environment, NC embedded into ceramic scaffolds and primed with osteogenic medium only sporadically formed intramembranous bone tissue. A clonal study could not demonstrate that the low bone formation efficiency was related to a possibly small proportion of cells competent to become fully functional osteoblasts. We next tested whether the cues present in an orthotopic environment could induce a more efficient direct osteoblastic transformation of NC. Using a nude rat calvarial defect model, we demonstrated that (i) NC directly participated in frank bone formation and (ii) the efficiency of survival and bone formation by NC was significantly higher than that of reference osteogenic cells, namely bone marrow-derived mesenchymal stromal cells. This study provides a proof-of-principle that NC have the plasticity to convert into bone cells and thereby represent an easily available cell source to be further investigated for craniofacial bone regeneration.

An improved cartilage digestion method for research and clinical applications.

Enzymatic isolation of chondrocytes from a cartilage biopsy is the first step to establish in vitro models of chondrogenesis or to generate cell-based grafts for cartilage repair. Such process is based on manually operated procedures and typically results in yields lower than 20% of the total available cells. In this study, we hypothesized that, as compared to conventionally used protocols, the enzymatic digestion of human articular cartilage in the presence of ascorbic acid 2-phosphate (AscA2P) or of sodium chloride (NaCl), in combination with the use of a perfusion bioreactor system, leads to a higher and more reproducible yield of cell populations with high proliferation and chondrogenic capacity. The addition of AscA2P within the enzymatic digestion medium did not significantly increase the cell yield, but resulted in a significant decrease of the intradonor variability in cell yield (-17.8% ± 10.7%, p = 0.0247) and in a significant increase of the proliferation rate of the isolated chondrocytes (+19.0% ± 1.4%, p < 0.05) with respect to the control group. The addition of NaCl during cartilage digestion did not modulate cell yield. When the cartilage digestion was further performed under direct perfusion flow, beneficial synergistic effects were achieved, with an overall increase of 34.7% ± 6.8% (p < 0.001) in the cell yield and an average decrease of 57.8% ± 11.2% (p < 0.01) in the coefficient of variation with respect to the control group. Importantly, by implementing this strategy it was possible to retrieve clonal subpopulations more efficiently capable of undergoing chondrogenesis, both in vitro and in vivo. Our findings bear relevance for the preparation of human chondrocytes for laboratory investigations, and in the perspective of efficient and streamlined manufacturing of cell/tissue grafts for articular cartilage repair.

Adult human neural crest-derived cells for articular cartilage repair.

In embryonic models and stem cell systems, mesenchymal cells derived from the neuroectoderm can be distinguished from mesoderm-derived cells by their Hox-negative profile--a phenotype associated with enhanced capacity of tissue regeneration. We investigated whether developmental origin and Hox negativity correlated with self-renewal and environmental plasticity also in differentiated cells from adults. Using hyaline cartilage as a model, we showed that adult human neuroectoderm-derived nasal chondrocytes (NCs) can be constitutively distinguished from mesoderm-derived articular chondrocytes (ACs) by lack of expression of specific HOX genes, including HOXC4 and HOXD8. In contrast to ACs, serially cloned NCs could be continuously reverted from differentiated to dedifferentiated states, conserving the ability to form cartilage tissue in vitro and in vivo. NCs could also be reprogrammed to stably express Hox genes typical of ACs upon implantation into goat articular cartilage defects, directly contributing to cartilage repair. Our findings identify previously unrecognized regenerative properties of HOX-negative differentiated neuroectoderm cells in adults, implying a role for NCs in the unmet clinical challenge of articular cartilage repair. An ongoing phase 1 clinical trial preliminarily indicated the safety and feasibility of autologous NC-based engineered tissues for the treatment of traumatic articular cartilage lesions.

Engineered autologous cartilage tissue for nasal reconstruction after tumour resection: an observational first-in-human trial.

BACKGROUND: Autologous native cartilage from the nasal septum, ear, or rib is the standard material for surgical reconstruction of the nasal alar lobule after two-layer excision of non-melanoma skin cancer. We assessed whether engineered autologous cartilage grafts allow safe and functional alar lobule restoration. METHODS: In a first-in-human trial, we recruited five patients at the University Hospital Basel (Basel, Switzerland). To be eligible, patients had to be aged at least 18 years and have a two-layer defect (≥50% size of alar subunit) after excision of non-melanoma skin cancer on the alar lobule. Chondrocytes (isolated from a 6 mm cartilage biopsy sample from the nasal septum harvested under local anaesthesia during collection of tumour biopsy sample) were expanded, seeded, and cultured with autologous serum onto collagen type I and type III membranes in the course of 4 weeks. The resulting engineered cartilage grafts (25 mm × 25 mm × 2 mm) were shaped intra-operatively and implanted after tumour excision under paramedian forehead or nasolabial flaps, as in standard reconstruction with native cartilage. During flap refinement after 6 months, we took biopsy samples of repair tissues and histologically analysed them. The primary outcomes were safety and feasibility of the procedure, assessed 12 months after reconstruction. At least 1 year after implantation, when reconstruction is typically stabilised, we assessed patient satisfaction and functional outcomes (alar cutaneous sensibility, structural stability, and respiratory flow rate). FINDINGS: Between Dec 13, 2010, and Feb 6, 2012, we enrolled two women and three men aged 76-88 years. All engineered grafts contained a mixed hyaline and fibrous cartilage matrix. 6 months after implantation, reconstructed tissues displayed fibromuscular fatty structures typical of the alar lobule. After 1 year, all patients were satisfied with the aesthetic and functional outcomes and no adverse events had been recorded. Cutaneous sensibility and structural stability of the reconstructed area were clinically satisfactory, with adequate respiratory function. INTERPRETATION: Autologous nasal cartilage tissues can be engineered and clinically used for functional restoration of alar lobules. Engineered cartilage should now be assessed for other challenging facial reconstructions. FUNDING: Foundation of the Department of Surgery, University Hospital Basel; and Krebsliga beider Basel.