Injury, Int. J.

Care Injured 41 (2010) 1172–1177

Contents lists available at ScienceDirect

Injury
journal homepage: www.elsevier.com/locate/injury

Treatment of chondral defects of the knee with one step matrix-assisted

technique enhanced by autologous concentrated bone marrow: In vitro
characterisation of mesenchymal stem cells from iliac crest and subchondral bone
Laura de Girolamo a, Giulia Bertolini b, Matteo Cervellin c, Gabriella Sozzi b, Piero Volpi c,*
a
Orthopaedic Biotechnologies Lab, IRCCS Galeazzi Orthopaedic Institute, Milan, Italy
b
Molecular-Cytogenetics Unit, Dept of Experimental Oncology, Istituto Nazionale Tumori, Milan, Italy
c
Sports Traumatology and Arthroscopic Unit, IRCCS Galeazzi Orthopaedic Institute, Milan, Italy

A R T I C L E I N F O A B S T R A C T

Keywords: Cartilage repair is still an unsolved problem. In the last years many cell-based treatments have been
Chondral lesion proposed, in order to obtain good regeneration of cartilage defects. The Autologous Matrix-Induced
Matrix-assisted technique Chondrogenesis technique (AMIC1) combines the micro-fracture procedure with the use of a specific
Bone marrow biological membrane.
Mesenchymal stem cells
The phenotypic feature of bone marrow cell population, harvested from iliac crest and knee
Chondrogenic potential
subchondral bone of patients treated with the AMIC1 technique, enhanced by autologous concentrated
bone marrow, was analysed to evaluate potential variations of the cell population. Samples of eleven
patients, with isolated chondral lesions grade III or IV were treated with the AMIC1 technique, enhanced
by the use of autologous concentrated bone marrow. A small fraction of bone marrow samples, both from
iliac crest and from the created micro-fractures, was analysed by FACS analysis and then cultured to
verify their proliferative and differentiation potential.
An average of 0.04% of concentrated bone marrow cells harvested from the iliac crest, presented
mesenchymal stem cell phenotype (CD34 /CD45low/CD271high), whereas just 0.02% of these cells were
identified from the samples harvested during the creation of micro-fractures at the knee. After two
passages in culture, cells expressed a peculiar profile for MSC. Only MSC from bone marrow could be
long-term propagated and were able to efficiently differentiate in the cultures. Although the AMIC1
approach has many advantages, the surgical technique in the application of the microfracture technique
remains essential and affects the final result.
ß 2010 Elsevier Ltd. All rights reserved.

Introduction (scaffolds) seeded with terminally differentiated cells or

Articular cartilage has a limited self-reparative capacity,
principally due to the absence of vasculature, which would allow
the progenitor cells to colonise the possible chondral lesion.20 In
the contemporary developed countries, the incidence of the
diagnosis of articular cartilage pathology is increasing, and as a
result, more cartilage repair procedures are being performed each
year. During the present decade, from 2000 to 2010, the attention
has been particularly directed to the research in order to obtain
better results in terms of prevention, diagnosis and/or treatment of
knee joint related problems.6,8
Promising results have been obtained with cell-based
therapies.10,19 These techniques involve the use of biomaterials
* Corresponding author.
E-mail address: volpi.piero@libero.it (P. Volpi).
progenitor cells. Although these new approaches have produced
encouraging in vitro data, none of these has yet proved to be able
to fully regenerate the articular cartilage tissue of the knee in
long-term clinical trials. Moreover, this kind of technique
involves two surgical stages, increasing patients’ discomfort,
risks, and costs.5,21
A very simple but efficient way of treating cartilage defects is
the ‘‘microfracture’’ technique.17 This approach, although leading
to good results,22 has known limitations as to the size of repairable
chondral lesions (up to 2 cm2). For larger defects, a matrix-based
bone marrow stimulating technique, called Autologous Matrix-
Induced Chondrogenesis (AMIC1) has been recently proposed. It
combines the ‘‘microfracture’’ technique with the use of a type I/III
porcine collagen matrix (Chondro-Gide1, Geistlich Pharma),
which is able to stabilise and protect the blood clot coming from
the subchondral bone. Although recently introduced, satisfactory
results have been already been reported.18,23

0020–1383/$ – see front matter ß 2010 Elsevier Ltd. All rights reserved.
doi:10.1016/j.injury.2010.09.027
 L. de Girolamo et al. / Injury, Int. J. Care Injured 41 (2010) 1172–1177
An important aspect of bone marrow stimulating techniques is
the real amount of mesenchymal stem cells (MSC) that derive from
the microfractured areas of the subchondral bone. A wide
variability between individual patients or surgeons, in terms of
presence of MSC has been reported. This variability affects
radically the clinical outcome.
Based on this observation, a prospective randomised clinical
study was performed to compare the results in patients treated
either with the standard AMIC1 technique, or with the AMIC1
technique combined with the use of autologous concentrated bone
marrow harvested from the iliac crest. The aim was to evaluate the
potential positive effect of increasing the number of progenitor cells
in repairing chondral lesions of the knee. The study is still in
progress, because some patients have not reached the time endpoint
of final follow up. The preliminary data on the first 15 patients of this
study are presented at minimum follow up of 6 months. At the same
time, an in vitro study has been performed, analysing the features of
the cell populations isolated from bone marrow both from the knee’s
subchondral bone and from the iliac crest, in order to reveal possible
quality and quantitative differences.
Material and methods
Surgical technique
Eleven patients, following their written informed consent, were
treated with the AMIC1 technique combined with use of
autologous concentrated bone marrow. Inclusion criteria were
age between 18 and 55 years, presence of one or two chondral
lesions type of III o IV according to the Outerbridge system,1 lesion
size between 2 and 8 cm2 with surrounding cartilage with lesions
less than type II per Outerbridge. Exclusion criteria were the
presence of more than two chondral defects, immuno-mediated
pathologies or knee infection including osteoarthritis, knee
instability, meniscus tears, malignancies, serious cardiologic
pathologies or problematic general conditions.
After arthroscopically confirming the indications to the treat-
ment, a small amount (24 ml) of bone marrow was harvested from
the iliac crest using a particular large bore needle. The sample was
then centrifuged for 15 min to obtain a concentrated phase
containing mononuclear cells, using the device MarrowStimTM
(Biomet Biotechnologies1). Meanwhile, a small incision was made
in correspondence to the localisation of the cartilage defect. The
[()TD$FIG]
Fig. 1. Concentrated bone marrow from the iliac crest after centrifugation (a) in which the cut membrane is dipped before be placed on the chondral defect (b).
1173
lesion was debrided in order to remove the soften cartilage and to
create a regular-shaped lesion. Using a template, the membrane was
then cut according to the dimensions and the size of the defect.
Following the Steadman’s technique,17 microfractures were then
created using specific awls (ChondropicTM, Arthrex1). According to
the locally developed technique, the membrane was then dipped
into the obtained concentrated bone marrow, and then glued as
usually (Fig. 1). The blood clot resulting from the microfractures was
stabilised and maintained placing the wet membrane on the defect
and fixing it with synthetic fibrin glue (Tissucol1, Baxter).
The rehabilitation program was different for the treated condylar
or patellar defects. For the condylar defect cases immediate full
range of motion was allowed without any weight bearing for 3
weeks. Full bearing was allowed after the sixth week. In the case of
patellar defects, patients were allowed to progressively restore the
full range of motion and bearing starting from the early post-
operative days. All cases were advised to return to sports after 4–6
months from surgery, depending of the type of sport.
Isolation, in vitro growth and characterisation of hMSCs
A small fraction of bone marrow samples retrieved both from
the iliac crest and from the microfracture sites, has been processed
in order to isolate, characterise and culture the mesenchymal stem
cells population.
The samples were washed with PBS and centrifuged twice at
900  g for 5 min. Cells were counted and plated at 2  105 cells/
cm2 in human Mesenchymal Stem Cells Growth Medium (MSCGM
– Lonza, Verviers, Belgium, control medium) in 75 cm2 flasks
(Corning Inc., NY, USA). After 72 h, non-adherent cells were
discarded and fresh medium was added to adherent cells. MSCGM
was replaced every 4 days. When reached the confluence, hMSCs
were detached by Trypsin/EDTA solution (Lonza) and then re-
seeded at 5–6  103 cells/cm2.
To determine the percentage of hMSCs in the bone marrow
aspirates, cell samples were assessed for the expression of specific
stem cell markers by cytofluorimetric analyses. Cells were stained
with conjugated antibodies CD271-PE (Miltenyi Biotech, Bergisch
Gladbach, Germany), CD45-FITC and with the vital dye 7-AAD (all
from Becton Dickinson, Heidelberg, Germany). Cells were re-
suspended in staining solution containing 1% BSA, 2 mM EDTA and
incubated for 30 min at 40 8C with specific antibodies at the
appropriate dilution and with 10 ml of vital dye 7-AAD.
1174 L. de Girolamo et al. / Injury, Int. J. Care Injured 41 (2010) 1172–1177
To characterise the phenotype of cultured hMSCs, cells grown for
two passages in control medium were stained with the following
conjugated antibodies: anti human CD45-PE, CD34-PE, CD31-PE,
CD90-PE, CD73-PE, CD166-PE, CD44-PE (all from Becton Dickinson)
and CD105-FITC (R&D System Minneapolis, MN, USA). Cells were
incubated with saturating concentrations of the appropriate
antibodies for 30 min at 40 8C and analysed by FACSCaliburTM
(Becton-Dickinson) and CellQuestTM analysis software.
In vitro differentiation assays
The differentiation potential of hMSCs was induced using
specific commercially available differentiation medium, all pur-
chased from Lonza. To obtain the chondrogenic differentiation,
5  105 hMSCs were re-suspended in Complete Chondrogenic
medium (supplemented with Dexamethasone, Ascorbate, ITS + -
Supplement, Sodium Pyruvate, Proline and 10 ng/ml TGFb3 –
Lonza), centrifuged at 150  g for 5 min. The resulting pellet was
left at the bottom of the tube in order to obtain a micromass made
of cells, and incubated at 37 8C in a humidified atmosphere of 5%
CO2, loosing the cap of the tube half-open to allow gas exchange.
Medium was replaced every 2–3 days for 3 weeks. The resulting
pellet was then formalin-fixed and paraffin embedded for
histological processing. Sections of 3 mm were slide-mounted,
stained with 1% Alcian blue solution, specific for sulphated
glycosaminoglycans, dissolved in 3% acetic acid pH 2, 5 for
30 min and counterstained with nuclear red solution.
For adipogenic differentiation, hMSCs were plated at a concen-
tration of 2  104 cells/cm2 in control medium. After 5 days
Adipogenesis Induction Medium (supplemented with Insulin, L-
glutamine, Dexamethasone, Indomethacin, 3-isobuty-l-methyl-
xanthine – Lonza) was added for 3 days, followed by 3 days of
[()TD$FIG]
culture in supplemented Adipogenic Maintenance Medium (Lonza).
Fig. 2. Cytogram distribution of bone marrow cell populations in iliac crest (left) and knee microfractures (right) of the same patients. A higher percentage of lymphoid and
myeloid progenitor cells (blast cells) have been observed in the sample harvested from iliac crest.
After 3 complete cycles of induction/maintenance, hMSCs were
maintained for 7 more days in Adipogenic Maintenance Medium,
replacing the medium every 2–3 days. Cells were then washed in
PBS, fixed with 10% formalin and stained with fresh Oil Red-O
solution for 10 min.
For osteogenic differentiation 3  103/cm2, hMSCs were plated in
control medium. After 24 h Osteogenesis Induction Medium
(supplemented with Ascorbate, L-glutamine, b-Glycerophosphate,
Dexamethasone – Lonza) was added to the plate, changing medium
twice a week for 21 days. To assay calcium deposition cells were
washed in PBS, fixed in a solution of ice-cold 70% ethanol and stained
with 1 ml of 2% Alizarin red S solution, pH 4.1 for 10 min.
Results
The mean age of the 11 patients (9 male, 2 female) was
30.3  10.5 (range 17–43). There were just one patient that had a
patellar chondral defect repair, the other were all treated for femoral
condyle defects. Since most of the enrolled patients have reached a
follow-up of just 6 months, the short term results could only be
reported.
No adverse events have been observed in these patients on both
groups for this period of time. No acute or long-lasting pain was
reported for any patients undergone the harvest of bone marrow
from iliac crest. The modified technique was reproducible and
quite simple, and allowed maintaining the same duration of
intervention with the classic AMIC1 technique.
An average of 0.04% of concentrated bone marrow cells
harvested from iliac crest presented mesenchymal stem cells
phenotype: CD34 /CD45low/CD271high. Just 0.02% of cells pre-
sented this expression pattern in the samples from the micro-
fractures blood samples (Fig. 2). After two passages in culture, cells
were analysed by FACS for the expression of mesenchymal cell
[()TD$FIG] L. de Girolamo et al. / Injury, Int. J. Care Injured 41 (2010) 1172–1177 1175

Fig. 3. Phenotypic profile of concentrated bone marrow MSC from iliac crest is after two passages in culture by cytofluorimetric analysis.
[()TD$FIG]
markers. The expression profile was peculiar for MSC: almost 100%
of the cells were positive for CD105, CD73, CD166 and CD90,
whereas no CD34 and CD45 positive cells were detected, excluding
a possible contamination of haematopoietic cells. In Fig. 3, a
representative cytofluorimetric analysis of passage II concentrated
bone marrow MSC from iliac crest is shown. Moreover, FSC analysis
revealed a significant difference in cytogram distribution between
knee microfracture blood and iliac crest samples: in particular the
percentage of lymphoid and myeloid progenitor cells (blast cells)
was 0.14 and 2.58, respectively.
To verify the multipotent differentiation of the isolated cells,
they were induced to differentiate into adipogenic, osteogenic and
chondrogenic lineage. Only MSC from bone marrow could be long-
term propagated and were able to efficiently differentiate, whereas
culture derived from microfracture can be propagated in vitro only
for few passages, indicating a limited self renewal potential. Cells
cultured in adipogenic medium for 28 days produced high level of Fig. 4. Adipogenic potential of MSC from concentrated bone marrow from iliac crest.
intracellular lipid drops, absent in undifferentiated cells, which Cells, after 28 days, in adipogenic medium, formed lipid droplets, which are stained
have been stained by Oil Red O (Fig. 4). by Oil Red O.
1176 [()TD$FIG]
L. de Girolamo et al. / Injury, Int. J. Care Injured 41 (2010) 1172–1177

To assess osteogenic differentiation, calcified matrix production

has been evaluated using Alizarin Red S. In undifferentiated cells
the staining was undetectable (Fig. 5a), whereas cells cultured for
21 days in osteogenic medium produced Alizarin Red-positive
calcified matrix on top of the cell monolayer, suggesting a
significant differentiation process (Fig. 5b).
Sections of MSC micromasses were evaluated to reveal the
presence of sulphated glycosaminoglycans. Only the micromasses
cultured for 21 days in chondrogenic medium were positive for
Alcian Blue staining, as shown in Fig. 6b, indicating an initial
chondrogenic differentiation process. Moreover, micromasses
maintained in non-inductive medium were smaller and less solid
than the ones cultured in chondrogenic medium, suggesting a poor
and not specific formation of extracellular matrix (Fig. 6a).

Discussion

Bone marrow stimulation techniques, besides enhancing

cytokines that promote cartilage repair, they also induce the
chondroprogenitors to migrate into the cartilage lesion.12
Currently, these techniques are very commonly used by the
majority of orthopaedic surgeons, as they are easy to perform,
need no particular surgical instruments and are less expensive
than others.4,9,16 However, as already reported,13 these methods
of repair are able to produce fibrocartilage tissue to the defects,
probably because the number of progenitors cells coming from
the bone marrow is too small to induce hyaline cartilage
formation.17
The AMIC1 technique can be included to the family of the
‘‘matrix-assisted techniques’’, which combine the regenerative
potential provided by a cell source (either undifferentiated or
terminally differentiated cells), with the effect of a specific matrix,
Fig. 6. Chondrogenic potential of MSC from concentrated bone marrow from iliac
able to induce the differentiation process. Differently from many crest. Sections of MSC micromasses (3 mm) cultured for 21 days in chondrogenic
others ‘‘matrix-assisted techniques’’, AMIC1 is performed in one
[()TD$FIG]
medium were positive for Alcian Blue staining (b), whereas not specific cartilaginous
matrix have been produced by cells maintained in non-inductive medium (a).

Fig. 5. Osteogenic potential of MSC from concentrated bone marrow from iliac crest.
Cells cultured for 21 days in osteogenic medium were able to differentiate
producing extracellular calcified matrix, positively stained by Alizarin Red S (b),
whereas cells maintained for the same time in non-inductive medium were not able
to produce it (a).
surgical step, thus reducing costs and discomfort for the patients.
The particular biochemical structure of the membrane (type I/III
porcine collagen) is able to create a suitable microenvironment to
induce progenitor cells, contained in the microfractures blood, and
acquire chondrocyte-like properties easier.
Since most of the patients in this series have just reached a follow
up of 6 months, and as this time point could be not indicative for the
regeneration chondral process, the evaluation of the clinical
outcome is still inconclusive. Similarly, other authors11,14 have
demonstrated that bone marrow stimulation techniques give very
good result in the short period, but then, at longer follow up, the
new-formed tissue start a degeneration process. However, recently
Gille et al.23 showed very good clinical results and significant
improvements respect to pre-operative values in term of Lysholm
Score, Tegner Activity Scale and ICRS score in patients treated with
the AMIC1technique at an average follow up of 24 and 36.
Additionally, at this short follow-up it is not possible to identify
potential differences between the two groups of patients included in
this study. Clinical evaluation is based on the calculation of the
Lysholm Knee Score, the IKDC score and the VAS pain scale, as
recorded preoperatively and at 6, 12 and 24 months from surgery. At
the same time point each patient will have an MRI of the treated
knee, in order to evaluate the filling of the cartilage defect.
Our preliminary data suggest that both types of treatment give
very good results after 6 months from surgery, with no significant
differences in term of patients’ satisfaction and clinical evaluation.
The modified technique does not increase the duration of the
intervention, because the centrifugation is performed during the
preparation of the chondral defects. Of course, the centrifugation,
used as a very simple way of bone marrow concentration, cannot
 L. de Girolamo et al. / Injury, Int. J. Care Injured 41 (2010) 1172–1177
produce a mass of purified MSC population, but, on the other hand, it
allows the completion of the procedure in one stage. No patients
reported long-lasting pain or subsequent complications related to
the harvest of bone marrow from iliac crest. The autologous use of
bone marrow avoids any potential risks from of disease transmission
or of reactions that allogenic grafts may cause.
The choice of embedding the membrane with concentrated
bone marrow allows the application of proportional quantities of
bone marrow to each individual chondral defect. According to this
technique developed locally, the membrane has been cut
according to the dimensions of the lesion, and then enriched
with the concentrated bone marrow, adjusting the quantity of the
absorbed cBM to the individual needs. Although it presents many
advantages, in comparison to the simple microfracture technique,
the AMIC1 approach provides a small quantity of progenitor cells
coming from the bone marrow, especially since the subchondral
bone at the site of the defect is pathological. Moreover, the
variability between different patients in terms of the presence,
viability and quantity of progenitor cells, complicate further the
standardisation of the results.
For these reasons, the evaluation of a probable positive effect of
enriching with greater numbers of progenitor cells the implant
covering chondral lesions of the knee appears important, and its
potentials appealing. The prospective study of the clinical outcome
of patients treated with the standard AMIC1 technique and with
the AMIC1 technique enhanced by the use of autologous
concentrated bone marrow harvested from iliac crest, as described
in this manuscript offers the means for this analysis. Our data
confirmed that the percentage of mesenchymal stem cells
contained in bone marrow from iliac crest is higher than the
one in marrow blood retrieved from subchondral bone. These cells
when cultured in vitro were able to efficiently proliferate and to
maintain their undifferentiated phenotype. Indeed, after two
passages in culture, they still expressed stem cell markers. Only
MSC isolated from iliac crest are able to actively proliferate for
many passages in culture and to efficiently differentiate. This
evidence, together with the observation of a higher number of
progenitor cells in bone marrow from iliac crest, seems to justify
our approach. The complete evaluation over a longer follow-up
period will be able to demonstrate the optimised outcome of
patients treated with the AMIC1 technique enhanced by autolo-
gous concentrated bone marrow.
In conclusion, although the ideal treatment for chondral defects
has not yet been identified, we strongly believe that the future
belongs to cell-based therapies.2,3,7,15 These procedures will have
to take into account, besides the efficacy, also the safety,
invasiveness, and costs related to them. Mesenchymal stem cells,
due to their availability and multipotent differentiation ability,
represent a promising cell source in the expanding field of
regenerative medicine.10
Role of the funding source
No study sponsor had a role in the study design; in collection,
analysis and interpretation of data; in the writing of the
1177
manuscript; in the decision to submit the manuscript for
publication.
Conflict of interest statement
No author of this paper has potential or actual conflict of
interest relevant to the subject matter or materials included in this
manuscript.
References
1. Cameron ML, Briggs KK, Steadman JR. Reproducibility and reliability of the
outerbridge classification for grading chondral lesions of the knee arthrosco-
pically. Am J Sports Med 2003;31:83–6.
2. Forriol F. Growth factors in cartilage and meniscus repair. Injury 2009;40(Suppl.
3):S12–6.
3. Gaissmaier C, Koh JL, Weise K. Growth and differentiation factors for cartilage
healing and repair. Injury 2008;39(Suppl. 1):S88–96.
4. Gaissmaier C, Koh JL, Weise K, Mollenhauer JA. Future perspectives of articular
cartilage repair. Injury 2008;39(Suppl. 1):S114–20.
5. Gates CB, Karthikeyan T, Fu F, Huard J. Regenerative medicine for the musculo-
skeletal system based on muscle-derived stem cells. J Am Acad Orthop Surg
2008;16:68–76.
6. Getgood A, Brooks R, Fortier L, Rushton N. Articular cartilage tissue engineer-
ing: today’s research, tomorrow’s practice? J Bone Joint Surg Br 2009;91:565–
76.
7. Giannoudis PV, Einhorn TA, Schmidmaier G, Marsh D. The diamond concept—
open questions. Injury 2008;39(Suppl. 2):S5–8.
8. Gobbi A, Bathan L. Biological approaches for cartilage repair. J Knee Surg
2009;22:36–44.
9. Hangody L, Vasarhelyi G, Hangody LR, et al. Autologous osteochondral graft-
ing—technique and long-term results. Injury 2008;39(Suppl. 1):S32–9.
10. Koga H, Engebretsen L, Brinchmann JE, et al. Mesenchymal stem cell-based
therapy for cartilage repair: a review. Knee Surg Sports Traumatol Arthrosc
2009;17:1289–97.
11. Kreuz PC, Steinwachs MR, Erggelet C, et al. Results after microfracture of full-
thickness chondral defects in different compartments in the knee. Osteoarthr
Cartilage 2006;14:1119–25.
12. Lotz M. Cytokines in cartilage injury and repair. Clin Orthop Relat Res
2001;S108–15.
13. Miller BS, Steadman JR, Briggs KK, et al. Patient satisfaction and outcome after
microfracture of the degenerative knee. J Knee Surg 2004;17:13–7.
14. Mithoefer K, McAdams T, Williams RJ, et al. Clinical efficacy of the microfracture
technique for articular cartilage repair in the knee: an evidence-based system-
atic analysis. Am J Sports Med 2009;37:2053–63.
15. Mollenhauer JA. Perspectives on articular cartilage biology and osteoarthritis.
Injury 2008;39(Suppl. 1):S5–12.
16. Pelttari K, Steck E, Richter W. The use of mesenchymal stem cells for chon-
drogenesis. Injury 2008;39(Suppl. 1):S58–65.
17. Steadman JR, Briggs KK, Rodrigo JJ, et al. Outcomes of microfracture for
traumatic chondral defects of the knee: average 11-year follow-up. Arthrosco-
py 2003;19:477–84.
18. Steinwachs MR, Kreuz PC, Guhlke-Steinwachs U, Niemeyer P. Current treat-
ment for cartilage damage in the patellofemoral joint. Orthopade 2008;37:841–
7.
19. Stoddart MJ, Grad S, Eglin D, Alini M. Cells and biomaterials in cartilage tissue
engineering. Regen Med 2009;4:81–98.
20. Vinatier C, Mrugala D, Jorgensen C, et al. Cartilage engineering: a crucial
combination of cells, biomaterials and biofactors. Trends Biotechnol
2009;27:307–14.
21. Willers C, Partsalis T, Zheng MH. Articular cartilage repair: procedures versus
products. Expert Rev Med Devices 2007;4:373–92.
22. Williams 3rd RJ, Harnly HW. Microfracture: indications, technique, and results.
Instr Course Lect 2007;56:419–28.
23. Gille J, Schuseil E, Wimmer J, et al. Mid-term results of Autologous Matrix-
Induced Chondrogenesis for treatment of focal cartilage defects in the knee.
Knee Surg Sports Traumatol Arthrosc 2010; Epub ahead of print.