PMID- 34403855
OWN - NLM
STAT- MEDLINE
DCOM- 20220406
LR  - 20220406
IS  - 1095-8673 (Electronic)
IS  - 0022-4804 (Linking)
VI  - 268
DP  - 2021 Dec
TI  - Histomorphometry in Peripheral Nerve Regeneration: Comparison of Different Axon 
      Counting Methods.
PG  - 354-362
LID - S0022-4804(21)00441-8 [pii]
LID - 10.1016/j.jss.2021.06.060 [doi]
AB  - BACKGROUND: Histomorphometry quantitatively evaluates nerve regeneration. Total 
      myelinated fiber count (TMFC) is most accurately obtained manually across full 
      nerve cross-sections, but most researchers opt for automated, sampled analysis. 
      Few of the numerous techniques available have been validated. The goal of this 
      study was to compare common histomorphometric methods (full manual [FM], sampled 
      manual [SM], and sampled automatic [SA]) to determine their reliability and 
      consistency. MATERIAL AND METHODS: Twenty-four rats underwent sciatic nerve (SN) 
      repair with 20mm isografts; SNs distal to the graft were analyzed. TMFC was 
      manually determined in each full cross-section. Counts were also extrapolated 
      from sampled fields, both manually and automatically with ImageJ software. 
      Myelinated fiber diameter, axon diameter, and myelin sheath thickness were 
      measured manually in full and sampled fields; G-ratio was calculated. 
      Repeated-measures MANOVA, Spearman correlation, and Wilcoxon signed-rank tests 
      were performed. A systematic review of histomorphometry in rat SN repair was 
      performed to analyze the variability of techniques in the literature. RESULTS: FM 
      TMFC was 13,506 +/- 4,217. Both sampled methods yielded significantly different 
      TMFCs (SM:14.4 +/- 13.4%, P< 0.001; SA:21.8 +/- 44.7%, P = 0.037). All three methods 
      strongly correlated with each other, especially FM and SM (r(s) = 0.912, P< 
      0.001). FM fiber diameter, axon diameter, and myelin sheath thickness did not 
      differ from SM (P = 0.493, 0.209, and 0.331, respectively). 65% of papers used 
      sampling; 78% utilized automated or semi-automated analysis. Software, sampling, 
      and histomorphometric parameters varied widely. CONCLUSION: SM and SA analysis 
      are reliable with standardized, systematic sampling. Transparency is essential to 
      allow comparison of data; meanwhile, researchers must be cognizant of the wide 
      variety of methodologies in the literature.
CI  - Copyright (c) 2021 Elsevier Inc. All rights reserved.
FAU - Orfahli, Lynn M
AU  - Orfahli LM
AD  - Department of Plastic Surgery, Cleveland Clinic Foundation, Cleveland, Ohio.
FAU - Rezaei, Majid
AU  - Rezaei M
AD  - Department of Plastic Surgery, Cleveland Clinic Foundation, Cleveland, Ohio.
FAU - Figueroa, Brian A
AU  - Figueroa BA
AD  - Department of Plastic Surgery, Cleveland Clinic Foundation, Cleveland, Ohio.
FAU - Crawford, Audrey V
AU  - Crawford AV
AD  - Department of Plastic Surgery, Cleveland Clinic Foundation, Cleveland, Ohio.
FAU - Annunziata, Michael J
AU  - Annunziata MJ
AD  - Department of Plastic Surgery, Cleveland Clinic Foundation, Cleveland, Ohio.
FAU - Rao, Maya
AU  - Rao M
AD  - Department of Plastic Surgery, Cleveland Clinic Foundation, Cleveland, Ohio.
FAU - Fahradyan, Vahe
AU  - Fahradyan V
AD  - Department of Plastic Surgery, Cleveland Clinic Foundation, Cleveland, Ohio.
FAU - Ordenana, Carlos X
AU  - Ordenana CX
AD  - Department of Plastic Surgery, Cleveland Clinic Foundation, Cleveland, Ohio.
FAU - Silver, Jerry
AU  - Silver J
AD  - Department of Neurosciences, Case Western Reserve University School of Medicine, 
      Cleveland, Ohio.
FAU - Rampazzo, Antonio
AU  - Rampazzo A
AD  - Department of Plastic Surgery, Cleveland Clinic Foundation, Cleveland, Ohio.
FAU - Gharb, Bahar Bassiri
AU  - Gharb BB
AD  - Department of Plastic Surgery, Cleveland Clinic Foundation, Cleveland, Ohio. 
      Electronic address: bassirb@ccf.org.
LA  - eng
PT  - Journal Article
PT  - Systematic Review
DEP - 20210814
PL  - United States
TA  - J Surg Res
JT  - The Journal of surgical research
JID - 0376340
SB  - IM
MH  - Animals
MH  - *Axons/physiology
MH  - Myelin Sheath/physiology
MH  - *Nerve Regeneration
MH  - Rats
MH  - Reproducibility of Results
MH  - Sciatic Nerve/surgery
OTO - NOTNLM
OT  - Autograft
OT  - Axon count
OT  - Histomorphometry
OT  - Isograft
OT  - Peripheral nerve regeneration
OT  - Rat sciatic nerve
COIS- Disclosure This research did not receive any specific grant from funding agencies 
      in the public, commercial, or not-for-profit sectors.
EDAT- 2021/08/18 06:00
MHDA- 2022/04/07 06:00
CRDT- 2021/08/17 20:17
PHST- 2020/11/20 00:00 [received]
PHST- 2021/04/16 00:00 [revised]
PHST- 2021/06/08 00:00 [accepted]
PHST- 2021/08/18 06:00 [pubmed]
PHST- 2022/04/07 06:00 [medline]
PHST- 2021/08/17 20:17 [entrez]
AID - S0022-4804(21)00441-8 [pii]
AID - 10.1016/j.jss.2021.06.060 [doi]
PST - ppublish
SO  - J Surg Res. 2021 Dec;268:354-362. doi: 10.1016/j.jss.2021.06.060. Epub 2021 Aug 
      14.