Verified Rigor RecordRigor Reviewed

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Injectable Genetic Engineering Hydrogel for Promoting Spatial Tolerance of Transplanted Kidney in Situ · 10.1002/advs.202408631

Name: Verified Rigor Record — Injectable Genetic Engineering Hydrogel for Promoting Spatial Tolerance of Transplanted Kidney in Situ · 10.1002/advs.202408631
Item: Injectable Genetic Engineering Hydrogel for Promoting Spatial Tolerance of Transplanted Kidney in Situ · 10.1002/advs.202408631
Rating: 75
Author: Αlpha¹

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Αlpha¹ Editorial

This rigor review was examined and confirmed by Αlpha¹ Editorial · July 6, 2026

Αlpha¹ Rigor Review

Rigor Reviewed

WARN

Warnings to address · Interventional Study

Eight dimensions — click a segment for details

Critical

High-priority

Model-reported confidence: highReviewer agreement: moderateEngine: 7.6.0Source: full text

Not verified in this report

No reported statistical tests were found to recompute.
No data or code availability links were detected to verify.

This Αlpha¹ Rigor Review uses AI Rigor Reviewers trained on a curated corpus of high-fidelity and retracted papers, with expert supervision and curation. It can still make mistakes; verify each finding against the source before relying on it.

The paper presents a well-motivated scientific premise and detailed methods, but has significant rigor and reporting gaps in study design (no randomization/blinding/power analysis), biological variables (mouse sex and housing not reported), key resources (missing catalog numbers/RRIDs), statistical reporting (no effect sizes or CIs), and a complete absence of data/code sharing. A few copyedit issues and an overstated conclusion ('transformative treatment') further reduce transparency. The study would benefit substantially from depositing raw data and addressing the missing rigor elements.

This assessment synthesizes three independent rigor evaluations, a copyedit pass, citation verification, and a claim audit. The statistics verification component found no test statistics with degrees of freedom or effect estimates with CIs to recompute, so mathematical plausibility is marked N/A. All eight dimensions were applicable and scored. The reviewers agreed substantially on the statuses, with minor divergence on study design (Reviewer 2 rated 'fail' vs. 'warn' from the other two) and ethical approvals (Reviewer 2 rated 'warn' vs. 'pass' from the others). My synthesis weighed the combined checklist evidence and scoring rules.

12 major claims checked against the paper's own evidence: 1 not fully backed by the presented evidence (unsupported or overstated).

Rigor

Eight Dimensions2 pass · 6 warn

12 major claims1 overstated1 partial10 supported

The introduction extensively cites prior research on immune tolerance strategies, engineered EVs, and hydrogels, and details limitations such as short in vivo half-life of cytokines and inability of EVs to form sustained tolerance niches. The premise that combining cytokine co-stimulation with contact-dependent molecules in a hydrogel can induce spatial tolerance is logically built from these cited limitations.

Sub-criteria

prior work citedADEQUATE

premise rationaleADEQUATE

limitations addressedADEQUATE

Per reviewer (3) — the votes behind the verdict

PASSReviewer 1· 90% conf

The paper clearly establishes the scientific premise, citing prior work on immune tolerance, regulatory T cells, extracellular vesicles, and hydrogels, and provides a logical rationale for the study objectives.

Evidence

paraphrase[Introduction, paragraphs 1-6]

“The introduction discusses limitations of existing cytokine-based and EV-based approaches, such as short half-lives and inability to form spatial tolerance niches.”

direct quote[Introduction, paragraph 6]

“Creating an independent immune tolerance space to facilitate cytokine co-stimulation and trigger contact-dependent molecular signaling thresholds is essential for altering the outcome of transplant rejection in kidney transplantation.”

direct quote[Introduction, paragraph 5]

“However, a noticeable limitation of EVs is their inability to maintain prolonged activity within the graft or to establish a tolerance space within the transplanted allograft.”

PASSReviewer 2· 90% conf

The scientific premise is well-established with adequate citation of prior work, a logical rationale, and acknowledgment of limitations of prior approaches.

Evidence

paraphrase[Introduction, paragraph 1-3]

“The introduction discusses the challenges of achieving co-inhibitory activation and cites previous studies on Treg-derived EVs and injectable hydrogels.”

paraphrase[Introduction, paragraph 4]

“It states that creating an independent immune tolerance space is essential for cytokine co-stimulation and triggering contact-dependent molecular thresholds.”

PASSReviewer 3· 90% conf

The paper establishes a clear scientific foundation for the need to create a tolerant immune niche, citing prior work on Treg-derived EVs, hydrogels, and transplant immunology.

Evidence

paraphrase[Introduction, paragraph 1–6]

“The introduction cites studies on immune tolerance, Treg-EVs, and injectable hydrogels, and identifies limitations such as short EV half-life and lack of spatial control.”

direct quote[Introduction, final paragraph]

“To address these challenges, we have designed an injectable, genetically engineered hydrogel, termed iGE‑Gel, in which genetic engineered Foe‑EVs were directly post‑inserted into the hydrogel network by the hydrophobic chains of HASA.”

direct quote[Discussion, paragraph 1]

“EVs have attracted considerable attention as drug delivery carriers… However, its short half‑life and off target effects severely hinder its clinical translation.”

This is an interventional animal study, so randomized allocation, blinding, power analysis, and inclusion/exclusion criteria are applicable. Randomization method and unit are not reported. Blinding of investigators/assessors is not mentioned. No a priori power/sample-size calculation is reported. Replicate distinction is addressed (n = 5 or 6 per group). Inclusion/exclusion criteria for animals are not pre-specified. Outlier handling is not described. Controls are adequate (Vector-Gel group, isograft group). Independent replication is not addressed (this is a single study).

Sub-criteria

controlsADEQUATE

power analysisNOT REPORTED

blinding levelsNOT REPORTED

outlier handlingNOT REPORTED

randomization unitNOT REPORTED

inclusion exclusionNOT REPORTED

randomization methodNOT REPORTED

replicate distinctionADEQUATE

independent replicationNOT REPORTED

Per reviewer (3) — the votes behind the verdict

WARNReviewer 1· 85% conf

The study design covers many applicable elements, but randomization and blinding are not reported, and no a priori power analysis is provided. Replicate definition is adequate, but inclusion/exclusion criteria and outlier handling are not described.

Evidence

absence[Methods, 'Mouse Model of Renal Transplant' and subsequent sections]

No randomization method is reported.

absence[Methods, 'Mouse Model of Renal Transplant' and 'Histology Staining']

No blinding is mentioned.

absence[Methods, 'Statistical Analysis']

No a priori power analysis.

direct quote[Figure legends (e.g., Figure 3, Figure 4)]

“n = 6, n = 5”

paraphrase[Results, 2.1 and Figures 3-6]

“Control groups include Vector-Gel (EVs from vector-transfected cells), free Foe-EVs, allograft, and isograft.”

FAILReviewer 2· 85% conf

The study design lacks several key rigor elements for an animal intervenational study: no randomization, blinding, power analysis, or explicit inclusion/exclusion criteria are reported.

Evidence

absence[Methods section]

No randomization method is described.

absence[Methods section]

No blinding is reported.

absence[Methods/Statistical Analysis]

No power analysis is reported.

WARNReviewer 3· 85% conf

The study includes key design controls (isograft, sham gel, free EV groups) and defines sample sizes, but lacks explicit randomization, blinding, a priori power analysis, and independent replication.

Evidence

direct quote[Section 2.3, paragraph 1 & Figure 3A]

“Balb/c (H‑2d) serving as the donor source and C57BL/6 (H‑2b) as the recipient, an alloreactive rejection model was established…”

direct quote[Experimental Section, Mouse Model of Renal Transplant]

“The mice, aged between 8–10 weeks and weighing 20–25 g, underwent anesthesia with 2% isoflurane.”

absence[Experimental Section]

No a priori power analysis is reported.

Mice are reported as 8-10 weeks old and 20-25 g, which is adequate for age/weight. However, the sex of the mice is not mentioned. Housing conditions (light cycle, temperature, enrichment) are not described. Species and strains (Balb/c, C57BL/6) are given, but vendor or RRID not provided.

Sub-criteria

demographicsN/A

sex reportedNOT REPORTED

sex justifiedN/A

age weight healthADEQUATE

housing conditionsNOT REPORTED

species strain sourceINADEQUATE

Per reviewer (3) — the votes behind the verdict

WARNReviewer 1· 80% conf

Mouse sex and age/weight are reported, and species and strain are clear, but housing conditions are not reported. For the porcine model, sex, age/weight, and source are reported, but housing conditions are not.

Evidence

direct quote[Methods, 'Mouse Model of Renal Transplant']

“The mice, aged between 8–10 weeks and weighing 20–25 g”

direct quote[Methods, 'Porcine Model of Renal Transplantation']

“The experiment was conducted using Bama male miniature pigs weighing 15–18 kg.”

absence[Methods, 'Mouse Model of Renal Transplant' and 'Porcine Model of Renal Transplantation']

Housing conditions are not described for either species.

WARNReviewer 2· 90% conf

Age and weight of mice are reported, but sex is not specified and housing conditions are not described.

Evidence

absence[Methods, Mouse Model section]

Sex of mice not reported.

direct quote[Methods, Mouse Model section]

“The mice, aged between 8–10 weeks and weighing 20–25 g, underwent anesthesia with 2% isoflurane.”

absence[Methods, Animal model section]

Housing conditions not described.

PASSReviewer 3· 90% conf

Biological variables (sex, age, weight, health status, species/strain, housing) are reported for mice and pigs.

Evidence

direct quote[Experimental Section, Mouse Model of Renal Transplant]

“Balb/c (H‑2d) donor mice provided kidneys for transplantation into C57BL/6 (H‑2b) recipient mice, following established procedures.”

direct quote[Experimental Section, Minimally Invasive Injection of iGE‑Gel under Laparoscopy]

“Male Bama miniature pigs were anesthetized by intramuscular injection…”

direct quote[Experimental Section, Mouse Model of Renal Transplant]

“The mice were then housed in a 28 °C incubator and closely monitored for vital signs for two days.”

The paper states that all animal protocols were approved by the Institutional Animal Care and Use Committee of the First Affiliated Hospital, Zhejiang University. This satisfies the key requirement for an animal study. Regulatory compliance is implied (use of aseptic technique, anesthesia, analgesia). No human subjects are involved.

Sub-criteria

iacuc statementADEQUATE

informed consentN/A

irb ethics statementN/A

regulatory complianceINADEQUATE

Per reviewer (3) — the votes behind the verdict

PASSReviewer 1· 90% conf

The paper reports approval from a named IACUC for the animal studies, which is adequate for an animal study. Regulatory compliance is implied but not explicitly stated in a single named framework.

Evidence

direct quote[Methods, 'Mouse Model of Renal Transplant']

“All animal protocols were approved by the Institutional Animal Care and Use Committee of the First Affiliated Hospital, Zhejiang University.”

absence[Methods, ethical statements]

No explicit statement of compliance with specific regulations like the Guide for the Care and Use of Laboratory Animals.

WARNReviewer 2· 85% conf

Animal ethics approval is stated but lacks a protocol number, and regulatory compliance is not explicitly mentioned.

Evidence

direct quote[Methods, Mouse Model section]

“All animal protocols were approved by the Institutional Animal Care and Use Committee of the First Affiliated Hospital, Zhejiang University.”

absence[Methods or Acknowledgments]

Regulatory compliance not stated.

PASSReviewer 3· 95% conf

Ethics approval is reported for the murine and porcine experiments with a named body (IACUC of the First Affiliated Hospital, Zhejiang University).

Evidence

direct quote[Experimental Section, Mouse Model of Renal Transplant]

“All animal protocols were approved by the Institutional Animal Care and Use Committee of the First Affiliated Hospital, Zhejiang University.”

absence[Experimental Section, Porcine Model of Renal Transplantation]

Regulatory compliance is not explicitly stated for the porcine model (e.g., Guide for the Care and Use of Lab Animals).

Antibodies are listed with supplier (Abcam) but without catalog numbers. Cell line HK-2 is used but source not specified. Mouse strains are named but no vendor or RRID. Reagents like stearic acid are described but no catalog number. Software versions are provided (FlowJo v10.8.1, GraphPad Prism v8.3.0) but no RRIDs. Mycoplasma testing is not reported.

Sub-criteria

mycoplasma testingNOT REPORTED

reagents identifiedINADEQUATE

organisms identifiedINADEQUATE

antibodies identifiedINADEQUATE

cell line authenticationNOT REPORTED

software tools identifiedINADEQUATE

Per reviewer (3) — the votes behind the verdict

WARNReviewer 1· 80% conf

Key biological and chemical resources are partially identified. The engineered cell line and EVs are described, and the hydrogel components (HASA, stearic acid, hyaluronic acid) are identified, but key reagents like antibodies and commercial kits lack catalog numbers or RRIDs, and software versions are missing.

Evidence

direct quote[Methods, 'Immunohistochemistry and Immunofluorescence']

“TNF-α (1:300, Abcam) or IL-1β (1:200, Abcam)”

direct quote[Methods, 'Statistical Analysis']

“GraphPad Prism v8.3.0”

absence[Methods, 'In Vitro Biocompatibility']

No cell line authentication or mycoplasma testing reported.

direct quote[Methods, 'Mouse Model of Renal Transplant', 'Porcine Model of Renal Transplantation']

“Balb/c (H-2d) donor mice provided kidneys for transplantation into C57BL/6 (H-2b) recipient mice”

WARNReviewer 2· 85% conf

Many key resources are mentioned but lack complete identification (e.g., catalog numbers, RRIDs, source details).

Evidence

direct quote[Methods, Immunofluorescence section]

“TNF-α (1:300, Abcam) or IL-1β (1:200, Abcam)”

absence[Methods, Biocompatibility section]

Cell line HK-2 source not provided.

direct quote[Methods, Statistical Analysis]

“FlowJo v10.8.1 software was utilized for data analysis. GraphPad Prism v8.3.0”

WARNReviewer 3· 85% conf

Key resources such as antibodies, reagents, and software are identified but lack catalog numbers, RRIDs, and version details.

Evidence

direct quote[Experimental Section, Immunohistochemistry and Immunofluorescence]

“primary antibodies targeting TNF‑α (1:300, Abcam) or IL‑1β (1:200, Abcam)”

direct quote[Experimental Section, Statistical Analysis]

“Statistical analysis was performed using the Statistical Package for the Social Sciences v22.0 and GraphPad Prism v8.3.0.”

absence[Experimental Section, In Vitro Biocompatibility]

Mycoplasma testing is not reported for the HK-2 cell line.

Tests used (Student's t-test, one-way ANOVA with Tukey post-hoc) are named. Software is identified. Error bars are defined as SEM and per-group n is often given. However, exact p-values are rarely reported; instead, thresholds (p<0.05, p<0.01, p<0.001) are used. Effect sizes and confidence intervals are not provided. Assumptions (normality, equal variance) are checked via Levene's and Kolmogorov-Smirnov tests, which is adequate.

Sub-criteria

tests namedADEQUATE

exact p valuesNOT REPORTED

effect sizes ciNOT REPORTED

data presentationADEQUATE

software identifiedADEQUATE

assumptions verifiedADEQUATE

mathematical plausibilityN/A

Per reviewer (3) — the votes behind the verdict

WARNReviewer 1· 75% conf

The statistical analysis has several strengths: tests are named, exact p-values are often reported, software is identified. However, test assumptions are not verified, effect sizes and confidence intervals are not systematically reported, and data presentation is mixed (bar charts with individual points sometimes not shown).

Evidence

direct quote[Methods, 'Statistical Analysis']

“One-way ANOVA was employed for normally distributed continuous variables, which was followed by a Tukey's post hoc test or a two-tailed Student's t-test.”

direct quote[Methods, 'Statistical Analysis']

“Prior to statistical analysis, Levene's test assessed variance homogeneity, while the normality of the data was evaluated using the Kolmogorov-Smirnov test.”

direct quote[Figure legends (e.g., Figure 3, Figure 4)]

“*** p < 0.001”

WARNReviewer 2· 85% conf

Statistical tests are named and software identified, but exact p-values and effect sizes are not reported; error bars are defined as SEM.

Evidence

direct quote[Throughout results]

“*** p < 0.001, compared to the allograft group”

direct quote[Methods, Statistical Analysis]

“Levene's test assessed variance homogeneity, while the normality of the data was evaluated using the Kolmogorov‐Smirnov test.”

WARNReviewer 3· 80% conf

Statistical tests are named (two-tailed t-test, one-way ANOVA with Tukey post-hoc, Mann-Whitney U), but assumptions verification is incomplete, exact p-values are often given as ranges (p < 0.001), and effect sizes with confidence intervals are not reported.

Evidence

direct quote[Experimental Section, Statistical Analysis]

“One‑way ANOVA was employed for normally distributed continuous variables, which was followed by a Tukey's post hoc test or a two‑tailed Student's t‑test. Conversely, the Mann‑Whitney U test was utilized to compare variables exhibiting normal distribution.”

direct quote[Figure 3 legend]

“*** p < 0.001, compared to the allograft group; ††† P < 0.001, compared to the Foe‑EVs group”

absence[Results and figures]

Effect sizes and confidence intervals are not reported for any comparison.

The paper does not contain any data availability statement, nor does it mention deposition of raw data (e.g., proteomics array data, flow cytometry data) in a repository. No code repository is provided. Given the computational nature of some analyses (flow cytometry, image analysis), this hinders reproducibility.

Sub-criteria

code sharingNOT REPORTED

accession numbersNOT REPORTED

repository depositNOT REPORTED

data availability statementNOT REPORTED

Per reviewer (3) — the votes behind the verdict

WARNReviewer 1· 80% conf

No data or code availability is mentioned. There is no statement about where the data or code can be accessed, which is a significant gap.

Evidence

absence[Entire paper]

No data availability statement.

absence[Entire paper]

No code sharing or repository deposits.

FAILReviewer 2· 90% conf

No data availability statement, repository deposit, or code sharing is provided.

Evidence

absence[Entire paper]

No data availability statement.

absence[Methods or Supporting Information]

No code sharing mentioned.

WARNReviewer 3· 90% conf

No data availability statement is present. Raw data are not deposited in a public repository. Code is not shared. This is a significant gap for reproducibility.

Evidence

absence[Entire paper]

No data availability statement is present anywhere in the manuscript.

absence[Entire paper]

No repository deposit or accession numbers are mentioned.

Methods are extensive (nearly all experimental procedures are described). Limitations are discussed in the Discussion section (short half-life of EVs, need for further optimization). Conclusions are generally proportional, claiming immunomodulation and survival benefit in murine models, which is supported by the data. Funding and COI: not explicitly stated in the provided text (may be in the acknowledgments section, which is not included). Trial registration and reporting guideline checklists are not applicable for this animal study. All outcomes: the paper reports extensive data, but no pre-specified outcome list is provided, making it difficult to assess if all outcomes are reported.

Sub-criteria

funding coiNOT REPORTED

trial registrationN/A

reporting guidelineNOT REPORTED

methods completenessADEQUATE

all outcomes reportedINADEQUATE

limitations discussedADEQUATE

conclusions proportionalADEQUATE

Per reviewer (3) — the votes behind the verdict

WARNReviewer 1· 85% conf

Methods are detailed and replicable, limitations are discussed, conclusions are proportional, and funding/COI is likely present (though not explicitly in the provided text). However, no trial registration or reporting guideline checklist is referenced, and it is not clear if all outcomes are reported.

Evidence

paraphrase[Methods, 'Preparation of Engineered EVs' through 'Statistical Analysis']

“All experimental methods are described in sufficient detail, including synthesis, cell culture, animal procedures, and statistical analysis.”

direct quote[Discussion, paragraphs 2,3,7]

“Future studies may focus on optimizing the composition and delivery schemes of iGE-Gel, exploring its potential in other transplantation models, and investigating the long-term effects and safety.”

absence[Entire paper (provided text)]

No funding or COI statement found.

WARNReviewer 2· 85% conf

Methods are detailed but lack some essential elements (blinding, randomization); no reporting guideline mentioned; funding and COI not stated.

Evidence

absence[Methods or Acknowledgments]

No reporting guideline mentioned.

absence[End of manuscript]

Funding and COI not stated.

WARNReviewer 3· 85% conf

Methods are detailed for most experiments, but the paper lacks a reporting checklist, trial registration, and a structured limitations section. Conclusions are generally proportional but occasionally overstate (e.g., 'transformative treatment').

Evidence

absence[Entire paper]

No ARRIVE or other reporting guideline is mentioned.

direct quote[Discussion, final paragraph]

“Future studies may focus on optimizing the composition and delivery schemes of iGE‑Gel, exploring its potential in other transplantation models, and investigating the long‑term effects and safety.”

direct quote[Abstract and Discussion]

“highlighting the potential of iGE‑Gel as a transformative treatment in allotransplant.”

14 copyedit issues flagged: mostly grammar, clarity, punctuation.

Checked 39 references: 1 verified — 38 not checked.

Αlpha¹ Rigor Action ItemsFinal pass before journal submission

This published paper has important transparency and rigor limitations that an informed reader should weigh carefully. The complete absence of a data availability statement and code sharing is a significant concern for reproducibility. The lack of randomization, blinding, and power analysis in the animal study design are methodological weaknesses that would warrant caution in interpreting causal claims. The overstatement 'transformative treatment' should be tempered. These issues collectively support the need for an erratum or independent re-analysis to verify the robustness of the reported findings.

1.
HIGHdata code
Add a data availability statement and deposit raw data (flow cytometry FCS files, ELISA plate reads, imaging files, survival data) in a public repository (e.g., Zenodo, Figshare, FlowRepository, GEO) with a persistent identifier. Update the manuscript to include a 'Data Availability' section with the repository link.
A data-driven preclinical study without any data availability or code sharing cannot be reproduced or verified, undermining the paper's credibility and violating journal transparency policies.
2.
HIGHrigor
Describe the method of randomization (e.g., computer-generated random numbers) and the unit of randomization (e.g., individual mouse) for the animal allocation in the 'Mouse Model of Renal Transplant' methods section.
Randomization is essential to avoid selection bias in interventional animal studies; its absence is a major design flaw flagged by all reviewers.
3.
HIGHrigor
State whether blinding was used during surgery, histology analysis, and flow cytometry data acquisition in the 'Mouse Model of Renal Transplant' and 'Statistical Analysis' sections; if not performed, provide a justification.
Blinding of investigators and assessors prevents bias in outcome measurement; its absence is a significant limitation that should be acknowledged.
4.
HIGHreporting
Temper the claim that iGE‑Gel is a 'transformative treatment' in the Abstract and Discussion, replacing it with language proportional to what a single preclinical study in one animal model can support (e.g., 'promising strategy' or 'potential immunomodulatory approach').
An overstated headline conclusion misrepresents the strength of the evidence and will attract reviewer criticism; the claim audit flagged this as an unsupported overstatement.
5.
HIGHethics
Add an explicit statement of compliance with animal welfare regulations (e.g., 'All procedures followed the Guide for the Care and Use of Laboratory Animals') and include the IACUC protocol number in the ethics section.
Regulatory compliance is expected by journals and reviewers; the current statement lacks a protocol number and does not reference specific guidelines.
6.
HIGHreporting
State the funding sources and grant numbers in a dedicated 'Funding' section, and include a conflict of interest statement even if the authors declare none.
Transparency about funding and competing interests is a standard reporting requirement; both are absent from the provided manuscript text.
7.
HIGHreporting
Include a completed ARRIVE 2.0 checklist or reference its use for the animal experiments in the Methods section.
ARRIVE guidelines are widely required by journals for animal studies; their absence is a notable reporting gap.
8.
HIGHstatistics
Report exact p-values (e.g., p = 0.023) instead of threshold asterisks for all primary comparisons, and include effect sizes (e.g., Cohen's d, mean difference) with 95% confidence intervals in figure legends and the text.
Threshold p-values limit the reader's ability to interpret the strength of evidence; effect sizes and CIs are needed for meaningful interpretation and meta-analysis.
9.
HIGHrigor
Provide an a priori power analysis or sample-size justification for the primary outcome (survival) in the 'Statistical Analysis' section, including the expected effect size, alpha, and power.
Without a power analysis, it is unclear whether the study had sufficient sample size to detect meaningful differences, a concern raised by all reviewers.
10.
MEDIUMother
Add catalog numbers and Research Resource Identifiers (RRIDs) for all primary antibodies listed in the 'Immunohistochemistry and Immunofluorescence' section.
Complete antibody identification is essential for reproducibility; catalog numbers and RRIDs are widely expected by journals.
11.
MEDIUMother
Report the source/vendor and authentication status (e.g., STR profiling) for the HK-2 cell line, and include a statement on mycoplasma testing in the 'In Vitro Biocompatibility' section.
Unidentified and untested cell lines risk cross-contamination or misidentification, compromising the validity of in vitro experiments.
12.
MEDIUMother
Explicitly state the sex of the mice used in the transplant model in the 'Mouse Model of Renal Transplant' section; if only one sex was used, justify why or acknowledge this as a limitation.
Sex is a critical biological variable; its omission hinders interpretation and generalizability of the findings.
13.
MEDIUMother
Describe housing conditions (light/dark cycle, temperature, humidity, enrichment) for both mice and pigs in the respective model subsections.
Housing conditions can affect physiology and behavior; their description is necessary for replication and is commonly requested by reviewers.
14.
MEDIUMother
Pre-specify the inclusion/exclusion criteria for animals (e.g., health checks, weight limits) and describe how outliers would be handled in the 'Mouse Model of Renal Transplant' and 'Statistical Analysis' sections.
Absent pre-specified criteria risk post hoc exclusions that could bias results; this was flagged by multiple reviewers.
15.
LOWcopyedit
Fix subject-verb agreement: change 'remain' to 'remains' in the Abstract sentence 'The establishment of a tolerant space… remain challenging'.
Minor grammar error that copyediting should correct for a polished submission.
16.
LOWcopyedit
In the Abstract, rephrase 'It is revealed that iGE-Gel displayed with abundant regulatory cytokines' to 'iGE-Gel displayed abundant regulatory cytokines' or 'It was revealed that iGE-Gel contains abundant regulatory cytokines'.
'Displayed with' is grammatically awkward and unclear.
17.
LOWcopyedit
In Figure 5 legend, insert a space before the double-dagger symbol (‡‡‡< 0.001 should be ‡‡‡ < 0.001) and ensure all symbols are explained in a single consistent notation.
Missing spaces and inconsistent symbol usage reduce readability and may confuse readers.

Full text (74,311 chars)Analyzed: Jul 6, 2026

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