Anders Lindahl - Academia.edu (original) (raw)

Papers by Anders Lindahl

PloS one, 2017

Cartilage repair and replacement is a major challenge in plastic reconstructive surgery. The deve... more Cartilage repair and replacement is a major challenge in plastic reconstructive surgery. The development of a process capable of creating a patient-specific cartilage framework would be a major breakthrough. Here, we described methods for creating human cartilage in vivo and quantitatively assessing the proliferative capacity and cartilage-formation ability in mono- and co-cultures of human chondrocytes and human mesenchymal stem cells in a three-dimensional (3D)-bioprinted hydrogel scaffold. The 3D-bioprinted constructs (5 × 5 × 1.2 mm) were produced using nanofibrillated cellulose and alginate in combination with human chondrocytes and human mesenchymal stem cells using a 3D-extrusion bioprinter. Immediately following bioprinting, the constructs were implanted subcutaneously on the back of 48 nude mice and explanted after 30 and 60 days, respectively, for morphological and immunohistochemical examination. During explantation, the constructs were easy to handle, and the majority ha...

Plastic and reconstructive surgery. Global open, 2017

The three-dimensional (3D) bioprinting technology allows creation of 3D constructs in a layer-by-... more The three-dimensional (3D) bioprinting technology allows creation of 3D constructs in a layer-by-layer fashion utilizing biologically relevant materials such as biopolymers and cells. The aim of this study is to investigate the use of 3D bioprinting in a clinically relevant setting to evaluate the potential of this technique for in vivo chondrogenesis. Thirty-six nude mice (Balb-C, female) received a 5- × 5- × 1-mm piece of bioprinted cell-laden nanofibrillated cellulose/alginate construct in a subcutaneous pocket. Four groups of printed constructs were used: (1) human (male) nasal chondrocytes (hNCs), (2) human (female) bone marrow-derived mesenchymal stem cells (hBMSCs), (3) coculture of hNCs and hBMSCs in a 20/80 ratio, and (4) Cell-free scaffolds (blank). After 14, 30, and 60 days, the scaffolds were harvested for histological, immunohistochemical, and mechanical analysis. The constructs had good mechanical properties and keep their structural integrity after 60 days of implanta...

Osteoarthritis and Cartilage, 2007

Purpose: Cell viability and matrix accumulation in scaffolds are important measurement outcomes o... more Purpose: Cell viability and matrix accumulation in scaffolds are important measurement outcomes of engineering cartilage tissues in vitro. Early assessments that predict longer-term outcomes would aid in more rapid screening of multiple scaffold-cell combinations. The objective of this study was to measure media metabolites for multiple chondrocyte-laden scaffolds at very early time points in culture, to evaluate their ability to predict extracellular matrix accumulation in the same samples at longer times. Methods and Materials: Porcine chondrocytes were encapsulated in 16 distinct formulations of a crosslinked elastin-like polypeptide (ELP). Constructs were cultured for 28 days, and culture media and pyruvate. On day 28, samples were assayed for accumulated sGAG. Regressions for accumulated sGAG on each metabolite were P126 Effect of different materials on the proliferation and migration of articular chondrocytes.

CARTILAGE, 2015

Objective An attempt to define pre-osteoarthritis (OA) versus early OA and definitive osteoarthri... more Objective An attempt to define pre-osteoarthritis (OA) versus early OA and definitive osteoarthritis. Methods A group of specialists in the field of cartilage science and treatment was formed to consider the nature of OA onset and its possible diagnosis. Results Late-stage OA, necessitating total joint replacement, is the end stage of a biological process, with many previous earlier stages. Early-stage OA has been defined and involves structural changes identified by arthroscopy or radiography. The group argued that before the “early-stage OA” there must exist a stage where cellular processes, due to the presence of risk factors, have kicked into action but have not yet resulted in structural changes. The group suggested that this stage could be called “pre-osteoarthritis” (pre-OA). Conclusions The group suggests that defining points of initiation for OA in the knee could be defined, for example, by traumatic episodes or surgical meniscectomy. Such events may set in motion metabolic...

Stem Cell and Gene-Based Therapy, 2006

Arthritis Research & Therapy

There is increasing interest in adeno-associated virus (AAV) vectors for a wide variety of gene t... more There is increasing interest in adeno-associated virus (AAV) vectors for a wide variety of gene therapy applications. AAV is a nonpathogenic human parvovirus that can mediate long-term transduction of a number of cell types without provoking a significant immune response. These properties make AAV especially attractive for use in gene therapy of rheumatoid arthritis (RA), a chronic inflammatory disease. To investigate the potential of AAV in gene therapy of arthritis, the ability of AAV to infect synovium in vitro and in vivo was tested. Three human RA synovial fibroblast cell lines and two murine (one DBA/1J and one DBA1J×C3H F1) synovial fibroblast cell lines were used to test AAV transduction in vitro. The cell lines (2 × 10 5 cells) were infected with 10 4 particles/cell of a murine IL-10-encoding vector (AAV-mIL-10) alone or with the addition of a low titer (100 particles/cell) of an E1-, E3-deleted recombinant adenovirus to provide E4orf6 activity to enhance second-strand synthesis. The supernatants were harvested from the wells at various time points and assayed for mIL-10 expression by ELISA. Both human synovial cell lines infected with AAV alone demonstrated low-level transgene expression throughout the course of the study. However, by day 10, all human cultures coinfected with adenovirus showed a 16-to 56-fold increase in mIL-10 compared to cultures infected with AAV-mIL10 alone. By day 30, a 31-to 135-fold increase was observed. No such increase was observed in any of the mouse cell lines. To determine the AAV transduction efficiency for synovium in vivo, human RA synovial tissues obtained from patients undergoing joint-replacement surgery were implanted subcutaneously on the backs of NOD.CB17-Prkdc SCID mice. After allowing a 2-week period for engraftment, tissues were injected with 3.4 × 10 11 particles of AAV-luciferase alone or in combination with 1.0 × 10 11 particles of adenovirus. Two weeks following AAV administration, the tissues were homogenized and assayed for expression of luciferase. Only the tissues coinfected with adenovirus had luciferase levels above background. A similar experiment with AAV-LacZ demonstrated X-gal staining only of synovial tissues coinfected with adenovirus. These findings demonstrate a preferential ability of AAV to transduce human, compared to mouse, synovial tissue and suggest that second strand synthesis may be a limiting factor in gene transduction. Further studies to elucidate the mechanisms limiting gene transduction in human synovium may allow optimization of this vector for the treatment of arthritis. P2 Delivery of antisense constructs and ribozymes to inhibit cartilage destruction in the SCID mouse model of RA

BioResearch Open Access, 2012

The standard culture system for in vitro cartilage research is based on cells in a three-dimensio... more The standard culture system for in vitro cartilage research is based on cells in a three-dimensional micromass culture and a defined medium containing the chondrogenic key growth factor, transforming growth factor (TGF)-b1. The aim of this study was to optimize the medium for chondrocyte micromass culture. Human chondrocytes were cultured in different media formulations, designed with a factorial design of experiments (DoE) approach and based on the standard medium for redifferentiation. The significant factors for the redifferentiation of the chondrocytes were determined and optimized in a two-step process through the use of response surface methodology. TGF-b1, dexamethasone, and glucose were significant factors for differentiating the chondrocytes. Compared to the standard medium, TGF-b1 was increased 30%, dexamethasone reduced 50%, and glucose increased 22%. The potency of the optimized medium was validated in a comparative study against the standard medium. The optimized medium resulted in micromass cultures with increased expression of genes important for the articular chondrocyte phenotype and in cultures with increased glycosaminoglycan/DNA content. Optimizing the standard medium with the efficient DoE method, a new medium that gave better redifferentiation for articular chondrocytes was determined.

Foot and Ankle Clinics, 2003

Cartilage and osteochondral lesions of the ankle are often results of a trauma and can cause grea... more Cartilage and osteochondral lesions of the ankle are often results of a trauma and can cause great problems for the patient. New clinical techniques have made it possible to diagnose and treat these injuries at an early stage and stop the deterioration that would otherwise likely occur. It is possible to detect early stage osteochondritis dissecans of the talus with MRI when the fragment is still in situ and not fragmented. Ankle arthroscopy is a useful tool for diagnosing intraarticular changes [1-4]. These lesions can be arthroscopically drilled or pinned with a successful result in patients with open epiphyses [1,2,5]; however, if the lesions are not diagnosed and treated early, this may lead to greater destruction of the lesion and thus present a clinical dilemma. A few different treatments have been proposed, such as debridement with drilling, microfracturing or bone grafting, osteochondral allo-and autografting (mosaic plasty), and autologous chondrocyte transplantation (ACT) [3,5]. A gold standard technique has not yet been established; the number of patients treated with these techniques is still too small, the time of follow-up is too short, or the results have not all been promising. At our clinic we prefer to use ACT. Our experience with ACT in the knee joint is that the treatment results in a repair tissue of hyaline-like characteristics mechanically, histologically and histochemically that corresponds well with a good and durable clinical outcome [6,7].

Joint Destruction in Arthritis and Osteoarthritis, 1993

Cellular aspects on articular cartilage growth and development are discussed. Cells with chondrog... more Cellular aspects on articular cartilage growth and development are discussed. Cells with chondrogenic potential are described and current treatment models for cartilage injuries are considered. A rabbit model for treatment of articular cartilage defects with autologous cultured and transplanted chondrocytes for treatment of knee cartilage defects in humans are discussed.

Comprehensive Physiology, 2011

The sections in this article are: 1 Studies Investigating the Validity of the Dual Effector Theor... more The sections in this article are: 1 Studies Investigating the Validity of the Dual Effector Theory 1.1 Differences Between Growth Hormone and Insulin-like Growth Factor I Actions In Vivo in Animals 1.2 Differences in Effects of Growth Hormone and Insulin-like Growth Factor I In Vivo in Humans 1.3 Localization of Growth Hormone Receptors: Indirect Evidence for a Local Effect of Growth Hormone 1.4 Local Action of Growth Hormone on Tissues 1.5 Effects of Growth Hormone and Insulin-like Growth Factor I on Cells of Different Maturation 1.6 Evidence In Vivo for a Stimulatory Effect of Growth Hormone, but not of Insulin-like Growth Factor I, on Progenitor Cells in the Growth Plate 2 Conclusion and Discussion 3 Modified Dual Effector Theory of Growth Hormone and Insulin-Like Growth Factor I Action in the Growth Plate

Pediatric Nephrology, 1991

A number of studies have shown that growth hormone (GH) and insulin-like growth factor-I (IGF-I) ... more A number of studies have shown that growth hormone (GH) and insulin-like growth factor-I (IGF-I) have important regulatory roles for skeletal growth. However, it has been a matter of controversy whether GH acts directly on cells in the growth plate or if the growth-promoting effects of GH are mediated by liver-derived (endocrine-acting) IGF-I. With the recognition that GH regulates the production of IGF-I in multiple extra-hepatic tissues, autocrine and paracrine functions of IGF-I have been suggested as important components of GH action. This review focuses on recent developments in our understanding of the cellular mechanisms by which GH promotes longitudinal bone growth and the inter-relationship between GH and IGF-I in the growth plate.

Osteoarthritis and Cartilage, 2006

via four G-protein-coupled receptors, EP1-EP4. We investigated role of PGE2 on mitochondrial func... more via four G-protein-coupled receptors, EP1-EP4. We investigated role of PGE2 on mitochondrial function and matrix degradation in chondrocytes. Methods: Cartilage explants were prepared from OA patients undergoing joint replacement surgery. In selected experiments chondrocytes were isolated using collagenase digestion and cultured in alginate beads. PGE2 receptor (EP1-4) expression was analyzed by FACS. Mitochondrial function was assessed by the fluorescent probe JC-1 and ATP generation. Results: Immunostaining for enzymes COX-1, COX-2, cPGES and mPGES confirmed the presence of PG biosynthetic pathway(s) in OA cartilage. All four PGE2 EP receptors were expressed in both normal and OA cartilage and confirmed in OA chondrocytes by FACS. However, the EP4 receptor was upregulated (2-3-fold) in OA compared to normal. Addition of PGE2 (0.1-10uM) exerted the following deleterious effects on OA chondrocytes: 1) decreased mitochondrial membrane activity (fluorescent probe JC-1) and ATP generation 2) inhibition of proteoglycan synthesis (35 S incorporation) by chondrocyte cultures to levels of 75% of control (p<0.01), 3) increased type II collagen degradation as assessed by C12C ELISA. 4) inhibited expression of aggrecan and type II collagen mRNA and 5) induced production of pro-MMP-13, IL-6 and IL-8, but inhibited MMP-1 secretion. The potency of PGE2 effects were comparable to that of IL-1b, and the inhibitory effects of a combination of IL-1 and PGE2 (10uM) were additive.To examine the effects of endogenous PGE2 produced in response to IL-1, we performed experiments in the presence or absence of the EP4 antagonist, A23858 or COX-2 selective inhibitor celecoxib. The addition of IL-1 reduced chondrocyte proteoglycan synthesis to 45% of control values; pretreatment with 10uM A23858 or 2uM celecoxib, reversed this inhibition to 67% and 65% respectively of control values (p<0.01). Both A23858 and celecoxib reversed other catabolic effects of IL-1 (1-4 above), including mitochondrial activity, ATP generation and inhibition of PGE2 dependent production of MMP-13. Conclusions: PGE2, the predominant eicosanoid produced by OA chondrocytes, exerts catabolic effects within cartilage. These deleterious effects are mediated, in part via the EP4 receptor, which should be considered as a potential target for disease modifying agents in OA.

Osteoarthritis and Cartilage, 2005

Objective: Clinical cartilage repair with transplantation of cultured chondrocytes, the first des... more Objective: Clinical cartilage repair with transplantation of cultured chondrocytes, the first described technique introduced in 1994, includes a periosteal membrane but today cells are also implanted without the periosteal combination. The aim of this study was to see if the periosteum had more than a biomechanical function and if the periosteum had a biological effect on the seeded cells tested in an agarose system in which the clonal growth in agarose and the external growth stimulation could be analysed. Methods: Four different experiments were used to study the growth of human chondrocytes in agarose and the periosteal influence. Human chondrocytes were isolated and transferred to either primary or secondary agarose culture. After 4 weeks, the total number of clones O 50 mm was counted. Cocultures of chondrocytes and periosteal tissue, cultures of chondrocytes with conditioned medium from chondrocytes, periosteal cells and fibroblast were used to study a potential stimulatory effect on growth and different cytokines and growth factors were analysed. Results: It was found that the human chondrocytes had different growth properties in agarose with the formation of four different types of clones: a homogenous clone without matrix production, a homogenous clone with matrix production, a differentiated clone with matrix production and finally a differentiated clone without matrix production. The periosteum exerted a paracrine effect on cultured chondrocytes in agarose resulting in a higher degree of cloning. The chondrocytes produced significant amounts of interleukin (IL)-6, IL-8, granulocyteemacrophage colony-stimulating factor (GM-CSF) and transforming growth factor (TGF)-b. The periosteum produced significant amounts of IL-6, IL-8 and TGF-b. Cocultures of chondrocytes and periosteum demonstrated a potentiation of IL-6 and IL-8 release but not of TGF-b and GM-CSF. Conclusion: Articular chondrocytes are able to form clones of different properties in agarose and the periosteum has a capacity of stimulating chondrocyte clonal growth and differentiation and secretes significant amounts of IL-6, IL-8, GM-CSF and TGF-b. It may be that the repair of cartilage defects with seeded chondrocytes could benefit from the combination with a periosteal graft. The production of TGF-b by implanted chondrocytes could influence the chondrogenic cells in the periosteum to start a periosteal chondrogenesis and together with the matrix from implanted chondrocyte production, a repair of cartilaginous appearance may develop; a dual chondrogenic response is possible.

Knee Surgery, Sports Traumatology, Arthroscopy, 2001

Knee Surgery, Sports Traumatology, Arthroscopy, 2010

The aim of our current study is to present the 12.6 years&amp;amp;amp;amp;amp;amp;amp;amp... more The aim of our current study is to present the 12.6 years&amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;#39; follow-up results in patients with cartilage lesions of the patellofemoral joint, treated with autologous chondrocyte implantation (ACI) with the use of periosteum. Ninety-two patients having patella or trochlea lesion participated in this study. Lysholm and Tegner questionnaires were completed 12.6 years (SD 2.3 years) after the surgery. The patients were asked whether they feel better, worse or had not experienced any difference compared to previous years and whether they would undergo the operation again. Complications or subsequent surgeries were also assessed. Median Tegner score was three, improved by one level compared with preoperative values (P = 0.02). Median Lysholm score was 70, improved by nine points (n.s.). Seventy-two percent of the patients were better or unchanged while 93% would undergo the operation again. Patients with no kissing lesions appeared to have a better prognosis. Patients with malalignment or instability that had undergone a realignment procedure had comparable outcomes to the patients that did not need any additional surgery. Realignment procedures increased the incidence of serious complications but they were associated with decreased incidence of periosteal hypertrophy. No association was found between the age of the patients at the time of the ACI or the size per lesion and any of the clinical outcomes. ACI provides a satisfactory outcome for the treatment of cartilage lesions of the patellofemoral joint, even for the cases with concomitant patellar instability. It seems that correcting the coexisting background factors with realignment, stabilizing or unloading procedures, along with the treatment of cartilage lesions, is improving the clinical outcomes over time and decreases the incidence of periosteal hypertrophies although increasing the incidence of serious complications. Our study reveals the good results and the high level of patients&amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;#39; activities (as shown by Tegner score), were preserved 12.6 years after the implantation, in both isolated trochlea and patella lesions and also in multiple and in kissing lesions where an intervention could be considered as a salvage procedure.

Knee Surgery, Sports Traumatology, Arthroscopy, 2012

The purpose of this study was to report on the clinical outcome of a large heterogenic cartilage ... more The purpose of this study was to report on the clinical outcome of a large heterogenic cartilage repair population treated with the profiling strategies of one experienced cartilage surgeon to provide evidence based tools for treatment selection in a clinical environment. Methods A total of 216 patients were identified in this prospective single-surgeon study. For the primary and secondary treatment of smaller defects, microfracture (MF) was used. Hyalograft C was used for first and second line larger defects, while carbon-fiber rod and pad implantations were used as a salvage procedure. Results Three years after the initial procedure, the clinical improvement was excellent for MF and Hyalograft C (P \ 0.001) and good for carbon-fiber procedures (P \ 0.05). Hyalograft C patients with prior anterior cruciate ligament reconstruction had less clinical improvement (P \ 0.05), while MF patients with prior cartilage repair were more likely to fail (Odds Ratio 20.5, P \ 0.05). Conclusion This is the first study that provides an assessment of the treatment strategies used by an experienced cartilage surgeon. A treatment algorithm for cartilage repair in a heterogenic population was created that based on the findings of this study could be implemented in a clinical environment. Level of evidence Prospective clinical case series, Level IV

Journal of molecular cell biology, 2010

Cardiotoxicity testing is a key activity in the pharmaceutical industry in order to detect detrim... more Cardiotoxicity testing is a key activity in the pharmaceutical industry in order to detect detrimental effects of new drugs. A reliable human in vitro model would both be beneficial in selection of lead compounds and be important for reducing animal experimentation. However, the human heart is a complex organ composed of many distinct types of cardiomyocytes, but cardiomyocyte clusters (CMCs) derived from human embryonic stem cells could be an option for a cellular model. Data on functional properties of CMCs demonstrate similarities to their in vivo analogues in human. However, development of an in vitro model requires a more thorough comparison of CMCs to human heart tissue. Therefore, we directly compared individually isolated CMCs to human fetal, neonatal, adult atrial and ventricular heart tissues. Real-time qPCR analysis of mRNA levels and protein staining of ion channels and cardiac markers showed in general a similar expression pattern in CMCs and human heart. Moreover, a si...

Clinical Orthopaedics and Related Research, 1996

Adult New Zealand rabbits were used to transplant autologously harvested and in vitro cultured ch... more Adult New Zealand rabbits were used to transplant autologously harvested and in vitro cultured chondrocytes into patellar chondral lesions that had been made previously and were 3 mm in diameter, extending down to the calcified zone. Healing of the defects was assessed by gross examination, light microscope, and histological-histochemical scoring at 8, 12, and 52 weeks. Chondrocyte transplantation significantly increased the amount of newly formed repair tissue compared to that found in control knees in which the lesion was solely covered by a periosteal flap. In another experiment, carbon fiber pads seeded with chondrocytes were used as scaffolds, and repair significantly increased at both 12 and 52 weeks compared to knees in which scaffolds without chondrocytes were implanted. The histologic quality scores of the repair tissue were significantly better in all knees in which defects were treated with chondrocytes compared to knees treated with periosteum alone and better at 52

Cells Tissues Organs, 2010

Notch signalling, via its downstream mediators HES1 and HES5, regulates development of several di... more Notch signalling, via its downstream mediators HES1 and HES5, regulates development of several different tissues. In vitro studies suggest that these genes are also involved in chondrogenesis and endochondral bone formation. In order to investigate the importance of HES1 and HES5 for these developmental processes, mice lacking chondrogenic expression of HES1 and HES5 were constructed by interbreeding HES5–/– mice homozygous for the floxed HES1 allele (HES1flox/flox) with COL2A1-Cre transgenic mice, creating conditional HES1;HES5 double mutant mice. The formation of cartilage and endochondral bone was studied in these mice using histological and immunohistochemical stainings, including Alcian Blue van Gieson, Safranin-O, modified Mallory Aniline Blue, tartrate-resistant acid phosphatase and collagen type II stainings. The mice were also studied using several different morphometrical analyses and the differentiation potential of the chondrocytes was evaluated in vitro. Unexpectedly, t...

Cells Tissues Organs, 2008

Background: Notch signalling controls differentiation and proliferation in various cell types and... more Background: Notch signalling controls differentiation and proliferation in various cell types and is associated with several diseases. We investigated the localization and regulation of several Notch markers in human osteoarthritic (OA) cartilage as well as identified genes controlled by Notch signalling. Methods: Immunolocalization and real-time PCR analysis of Notch markers in healthy and OA articular cartilage were performed. Genes regulated by Notch signalling were studied using microarray. Cytokine-induced transcription of Notch markers was analyzed using real-time PCR and its effect on cellular localization of the intracellular domain of Notch1 (NICD1) was investigated using immunohistochemistry, subcellular fractionation, and transfection. The effect of NFĸB activation on HES5 transcription was studied using the NFĸB inhibitor pyrrolidine dithiocarbamate. Results: Notch signalling was activated in OA cartilage and Notch1, Jagged1, and HES5 were abundantly expressed compared t...